Ginzberg D


Full name : Ginzberg Dalia

First name : Dalia

Mail : Dept. of Biol. Chem., The Institute of Life Sciences, The Hebrew University, 91904 Jerusalem

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Country : Israel

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Phone : (972) 2 585450

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References (25)

Title : Muscarinic modulations of neuronal anticholinesterase responses - Salmon_2005_Chem.Biol.Interact_157-158_105
Author(s) : Salmon A , Erb C , Meshorer E , Ginzberg D , Adani Y , Rabinovitz I , Amitai G , Soreq H
Ref : Chemico-Biological Interactions , 157-158 :105 , 2005
Abstract : Anticholinesterases (antiChEs) are increasingly used for treating patients with neurodegenerative diseases, but the dependence of their effects on the integrity of cholinergic functions has not yet been analyzed at the molecular level. Here, we report that manipulation of muscarinic neurotransmission confers drastic changes on antiChE responses in the rat brain. In the brains of naive, un-stressed rats, the irreversible organophosphate antiChE, diisopropylfluorophosphonate (DFP) induced post-treatment accumulation of catalytically active G1 monomers of acetylcholinesterase (AChE). Pre-treatment with the selective M1 muscarinic antagonist, pirenzepine, but not the general muscarinic antagonist, scopolamine, attenuated this G1 increase. DFP-enhanced AChE gene expression was accompanied by diverted splicing from the primary AChE-S mRNA variant, encoding G4 synaptic membrane AChE-S tetramers, to "readthrough" AChE-R mRNA, which encodes soluble G1 monomers. Both the mRNA increase and the shifted splicing were long lasting (>24 h) and common to the parietal cortex and hippocampal CA1 and CA3 neurons. Importantly, the splicing shift was maximal under DFP alone, as compared with sham-injected rats, and virtually preventable by pre-treatment with pirenzepine. In contrast, induction of AChE transcription was less dependent on muscarinic function, resulting in AChE-S but not AChE-R increases. Our findings demonstrate distinct regulation of the enhanced transcription and the alternative splicing reactions to antiChE treatment and shed new light on the differential responses to antiChEs of demented patients with increasingly impaired cholinergic neurotransmission.
ESTHER : Salmon_2005_Chem.Biol.Interact_157-158_105
PubMedSearch : Salmon_2005_Chem.Biol.Interact_157-158_105
PubMedID: 16289123

Title : The molecular neurobiology of acetylcholinesterase variants. -
Author(s) : Meshorer E , Cohen O , Yirmya R , Ginzberg D , Glick D
Ref : Cholinergic Mechanisms, CRC Press :119 , 2004

Title : Endotoxin-induced changes in human working and declarative memory associate with cleavage of plasma readthrough acetylcholinesterase - Cohen_2003_J.Mol.Neurosci_21_199
Author(s) : Cohen O , Reichenberg A , Perry C , Ginzberg D , Pollmacher T , Soreq H , Yirmiya R
Ref : Journal of Molecular Neuroscience , 21 :199 , 2003
Abstract : Endotoxin stimulation of the immune system produces marked alterations in memory functioning. However, molecular links between this cognitive response and infection-responding neurotransmission pathways are still unknown. The cytokine and memory responses of volunteers injected with 0.8 ng/kg Salmonella endotoxin were compared with changes in plasma levels and integrity of the stress-induced acetylcholinesterase variant, AChE-R. Vascular endothelial cells were found to express AChE-R messenger RNA and protein both in healthy and inflamed human tissues. Plasma AChE activity was reduced after endotoxin treatment, but not placebo treatment, parallel to the decline in cortisol after the endotoxin-induced peak and inversely to the accumulation of a C-terminal immunopositive AChE-R peptide of 36 amino acid residues. AChE-R cleavage coincided with significant endotoxin-induced improvement in working memory and impairment in declarative memory. By 3 h posttreatment, working memory improvement was negatively correlated with AChE-R cleavage, which showed association to proinflammatory cytokine levels. By 9 h posttreatment, declarative memory impairment was negatively correlated with AChE-R cleavage and positively correlated with the suppressed AChE activity. Endotoxin-induced peripheral cholinergic stress responses are hence associated with greater impairment in declarative memory and lower improvement in working memory, pointing at AChE-R as a surrogate marker of psychoneuroimmunological stress.
ESTHER : Cohen_2003_J.Mol.Neurosci_21_199
PubMedSearch : Cohen_2003_J.Mol.Neurosci_21_199
PubMedID: 14645987

Title : Neuronal overexpression of 'readthrough' acetylcholinesterase is associated with antisense-suppressible behavioral impairments - Cohen_2002_Mol.Psychiatry_7_874
Author(s) : Cohen O , Erb C , Ginzberg D , Pollak Y , Seidman S , Shoham S , Yirmiya R , Soreq H
Ref : Mol Psychiatry , 7 :874 , 2002
Abstract : Molecular origin(s) of the diverse behavioral responses to anticholinesterases were explored in behaviorally impaired transgenic (Tg) FVB/N mice expressing synaptic human acetylcholinesterase (hAChE-S). Untreated hAChE-S Tg, unlike nave FVB/N mice, presented variably intense neuronal overexpression of the alternatively spliced, stress-induced mouse "readthrough" mAChE-R mRNA. Both strains displayed similar diurnal patterns of locomotor activity that were impaired 3 days after a day-to-night switch. However, hAChE-S Tg, but not FVB/N mice responded to the circadian switch with irregular, diverse bursts of increased locomotor activity. In social recognition tests, controls displayed short-term recognition, reflected by decreased exploration of a familiar, compared to a novel juvenile conspecific as well as inverse correlation between social recognition and cortical and hippocampal AChE specific activities. In contrast, transgenics presented poor recognition, retrievable by tetrahydroaminoacridine (tacrine, 1.5 mg kg(-1)). Tacrine's effect was short-lived (24 h) suppression of the abnormal social recognition pattern in transgenics. Efficacy of antisense treatment was directly correlated with AChE-R levels and the severity of the impaired phenotype, being most apparent in transgenics presenting highly abnormal pre-treatment behavior. These findings demonstrate that neuronal AChE-R overproduction is involved in various behavioral impairments and anticholinesterase responses, and point to the antisense strategy as a potential approach for re-establishing cholinergic balance.
ESTHER : Cohen_2002_Mol.Psychiatry_7_874
PubMedSearch : Cohen_2002_Mol.Psychiatry_7_874
PubMedID: 12232781

Title : Antisense prevention of neuronal damages following head injury in mice - Shohami_2000_J.Mol.Med.(Berl)_78_228
Author(s) : Shohami E , Kaufer D , Chen Y , Seidman S , Cohen O , Ginzberg D , Melamed-Book N , Yirmiya R , Soreq H
Ref : J Mol Med (Berl) , 78 :228 , 2000
Abstract : Closed head injury (CHI) is an important cause of death among young adults and a prominent risk factor for nonfamilial Alzheimer's disease. Emergency intervention following CHI should therefore strive to improve survival, promote recovery, and prevent delayed neuropathologies. We employed high-resolution nonradioactive in situ hybridization to determine whether a single intracerebro-ventricular injection of 500 ng 2'-O-methyl RNA-capped antisense oligonucleotide (AS-ODN) against acetylcholinesterase (AChE) mRNA blocks overexpression of the stress-related readthrough AChE (AChE-R) mRNA splicing variant in head-injured mice. Silver-based Golgi staining revealed pronounced dendrite outgrowth in somatosensory cortex of traumatized mice 14 days postinjury that was associated with sites of AChE-R mRNA overexpression and suppressed by anti-AChE AS-ODNs. Furthermore, antisense treatment reduced the number of dead CA3 hippocampal neurons in injured mice, and facilitated neurological recovery as determined by performance in tests of neuromotor coordination. In trauma-sensitive transgenic mice overproducing AChE, antisense treatment reduced mortality from 50% to 20%, similar to that displayed by head-injured control mice. These findings demonstrate the potential of antisense therapeutics in treating acute injury, and suggest antisense prevention of AChE-R overproduction to mitigate the detrimental consequences of various traumatic brain insults.
ESTHER : Shohami_2000_J.Mol.Med.(Berl)_78_228
PubMedSearch : Shohami_2000_J.Mol.Med.(Berl)_78_228
PubMedID: 10933585

Title : Transgenic mammary gland expression of readthrough human AChE: A model system for cholinesterase regulation in mammalian body fluids -
Author(s) : Salmon AY , Sternfeld M , Ginzberg D , Patrick J , Soreq H
Ref : Journal de Physiologie (Paris) , 92 :489 , 1998

Title : Multilevel Approaches to AChE-Induced Impairments in Learning and Memory -
Author(s) : Seidman S , Cohen O , Ginzberg D , Eichengreen E , Pollak Y , Yirmiya R , Soreq H
Ref : In: Structure and Function of Cholinesterases and Related Proteins - Proceedings of Sixth International Meeting on Cholinesterases , (Doctor, B.P., Taylor, P., Quinn, D.M., Rotundo, R.L., Gentry, M.K. Eds) Plenum Publishing Corp. :183 , 1998

Title : In vitro phosphorylation of acetylcholinesterase at non-consensus protein kinase A sites enhances the rate of acetylcholine hydrolysis - Grifman_1997_Brain.Res.Mol.Brain.Res_51_179
Author(s) : Grifman M , Arbel A , Ginzberg D , Glick D , Elgavish S , Shaanan B , Soreq H
Ref : Brain Research Mol Brain Res , 51 :179 , 1997
Abstract : Here, we report that the catalytic subunit of cAMP-dependent protein kinase (PKA) but not casein kinase II or protein kinase C phosphorylates recombinant human acetylcholinesterase (AChE) in vitro. This enhances acetylthiocholine hydrolysis up to 10-fold as compared to untreated AChE, while leaving unaffected the enzyme's affinity for this substrate and for various active and peripheral site inhibitors. Alkaline phosphatase treatment enhanced the electrophoretic migration, under denaturing conditions, of part of the AChE proteins isolated from various mammalian sources and raised the isoelectric point of some of the treated AChE molecules, indicating that part of the AChE molecules are also phosphorylated in vivo. Enhancement of acetylthiocholine hydrolysis also occurred with Torpedo AChE, which has no consensus motif for PKA phosphorylation. Further, mutating the single PKA site in human AChE (threonine-249) did not prevent this enhancement, suggesting that in both cases it was due to phosphorylation at non-consensus sites. In vivo suppression of the acetylcholine hydrolyzing activity of AChE and consequent impairment in cholinergic neurotransmission occur under exposure to both natural and pharmacological compounds, including organophosphate and carbamate insecticides and chemical warfare agents. Phosphorylation of AChE may possibly offer a rapid feedback mechanism that can compensate for impairments in cholinergic neurotransmission, modulating the hydrolytic activity of this enzyme and enabling acetylcholine hydrolysis to proceed under such challenges.
ESTHER : Grifman_1997_Brain.Res.Mol.Brain.Res_51_179
PubMedSearch : Grifman_1997_Brain.Res.Mol.Brain.Res_51_179
PubMedID: 9427520

Title : Genetic manipulations of cholinergic communication reveal trans-acting control mechanisms over acetylcholine receptors. - Broide_1997_J.Recept.Signal.Transduct.Res_17_279
Author(s) : Broide RS , Grifman M , Shapira M , Ginzberg D , Soreq H
Ref : Journal of Receptor & Signal Transduction Research , 17 :279 , 1997
Abstract : Several approaches have been developed for genetic modulations of receptor expression. These initiated with gene cloning and heterologous expression in microinjected Xenopus oocytes, and proceeded through transgenic expression and genomic disruption of receptor genes in mice. In addition, antisense treatments have reduced receptor levels in a transient, reversible manner. Integration of foreign DNA with host genomic sequences yields both cis- and trans-acting responses. These may depend on the DNA integration site, host cells condition and most importantly, the affected signal transduction circuit. For example, acetylcholinesterase (AChE) overexpression in microinjected Xenopus tadpoles has been shown to upregulate alpha-bungarotoxin binding levels, indicating trans-acting control conferring overproduction of muscle nicotinic acetylcholine receptors. In transgenic mice expressing human AChE, the hypothermic response to oxotremorine was suppressed, reflecting modified levels of brain muscarinic receptors. To dissociate the feedback processes occurring in transfected cells from responses related to DNA integration, we examined the endogenous expression of the alpha 7 neuronal nicotinic acetylcholine receptor in PC12 cells transfected with DNA vectors carrying alternative splicing variants of human AChE mRNA. Our findings demonstrate suppression of alpha 7 receptor levels associated with the accumulation of foreign DNA in the transfected cells. Acetylcholine receptor levels thus depend on multiple elements, each of which should be considered when genetic interventions are employed.
ESTHER : Broide_1997_J.Recept.Signal.Transduct.Res_17_279
PubMedSearch : Broide_1997_J.Recept.Signal.Transduct.Res_17_279
PubMedID: 9029496

Title : Overexpression of alternative human acetylcholinesterase forms modulates process extensions in cultured glioma cells - Karpel_1996_J.Neurochem_66_114
Author(s) : Karpel R , Sternfeld M , Ginzberg D , Guhl E , Graessmann A , Soreq H
Ref : Journal of Neurochemistry , 66 :114 , 1996
Abstract : In addition to its well-known synaptic function, acetylcholinesterase was recently shown to stimulate neurite outgrowth from cultured chick neurons in a manner unrelated to its catalytic activity. It remained unclear, however, whether each of the variant acetylcholinesterase enzyme forms can promote such process extension and whether this effect of acetylcholinesterase was limited to neurite outgrowth. Using DNA microinjections and stable transfections of cultured glioma cells, we explored the possibility that specific acetylcholinesterase isoforms affect cellular development and morphology of CNS astrocytes. Cells microinjected with human ACHEDNA constructs that differ in their exon-intron composition displayed rapid yet stable induction of cell body enlargement and process extensions. Cells transfected with ACHEDNA carrying the neuronal-characteristic 3'-E6 domain also displayed stable process extensions. However, stable transfections with ACHEDNAs including the 3'-alternative 14/E5 region induced the appearance of small, round cells in a dominant manner. This was associated with expression of 14/E5-ACHEmRNA transcripts and the production of soluble acetylcholinesterase monomers that were catalytically indistinguishable from the 3'-E6 enzyme but displayed higher electrophoretic mobility than that of the 3'-E6 form. Thus, variable expression levels and alternative splicing modes of the ACHE gene correlated in these experiments with glial development in a manner that was apparently unrelated to catalysis.
ESTHER : Karpel_1996_J.Neurochem_66_114
PubMedSearch : Karpel_1996_J.Neurochem_66_114
PubMedID: 8522942

Title : Antisense Oligonucleotides Suppressing Expression of Cholinesterase Genes Modulate Hematopoiesis in Vivo and ex Vivo -
Author(s) : Soreq H , Lev-Lehman E , Patinkin D , Grifman M , Ehrlich G , Ginzberg D , Eckstein F , Zakut H
Ref : In Enzyme of the Cholinesterase Family - Proceedings of Fifth International Meeting on Cholinesterases , (Quinn, D.M., Balasubramanian, A.S., Doctor, B.P., Taylor, P., Eds) Plenum Publishing Corp. :1 , 1995

Title : Antisense inhibition of acetylcholinesterase gene expression causes transient hematopoietic alterations in vivo - Lev-Lehman_1994_Gene.Ther_1_127
Author(s) : Lev-Lehman E , Ginzberg D , Hornreich G , Ehrlich G , Meshorer A , Eckstein F , Soreq H , Zakut H
Ref : Gene Therapy , 1 :127 , 1994
Abstract : Hematopoietic acetylcholinesterase (ACHE) gene expression and its implication for development were studied by in vivo administration to mice of an antisense phosphorothioate oligonucleotide targetted toward ACHE (AS-ACHE). Hematopoietic alterations were observed by differential cell counts and ACHE mRNA levels determined by quantified RNA polymerase chain reaction (RNA-PCR) and in situ hybridization analyses. In control mice, injected with phosphate-buffered saline and untreated, ACHE mRNA labeling with ACHE [35S]cRNA was about 10-fold higher on megakaryocytes (MK) compared with all other bone marrow cells and increased by 20-fold during MK development, similar to reports for MK actin mRNA. Drastic reductions occurred in the bone marrow lymphocyte and erythroid fractions 12 days following intraperitoneal injection of AS-ACHE (5 micrograms/g weight) into groups of four mice. RNA-PCR revealed over 1000-fold decreases in ACHE mRNA levels in lymph nodes and bone marrow at this time, while actin mRNA levels dropped by 10 and 100-fold in lymph nodes and bone marrow of AS-ACHE treated mice compared with controls. In view of the developmental increase in MK actin, this suggested arrest in MK development as well. By 20 days postinjection, bone marrow actin mRNA was fully restored and the sensitive in situ hybridization technique revealed that ACHE mRNA levels were also restored and reached levels only 2-3-fold lower than in controls in all bone marrow cells of AS-ACHE treated mice. Moreover, lymphocytes and erythroid cells repopulated to levels 25% above normal, and promegakaryocyte and mature MK fractions of the total MK were 3 and 2-fold higher, respectively, than in controls.
ESTHER : Lev-Lehman_1994_Gene.Ther_1_127
PubMedSearch : Lev-Lehman_1994_Gene.Ther_1_127
PubMedID: 7584068

Title : Antisense oligonucleotide inhibition of acetylcholinesterase gene expression induces progenitor cell expansion and suppresses hematopoietic apoptosis ex vivo - Soreq_1994_Proc.Natl.Acad.Sci.U.S.A_91_7907
Author(s) : Soreq H , Patinkin D , Lev-Lehman E , Grifman M , Ginzberg D , Eckstein F , Zakut H
Ref : Proc Natl Acad Sci U S A , 91 :7907 , 1994
Abstract : To examine the role of acetylcholinesterase (EC in hematopoietic cell proliferation and differentiation, we administered a 15-mer phosphorothioate oligonucleotide, antisense to the corresponding ACHE gene (AS-ACHE), to primary mouse bone marrow cultures. Within 2 hr of AS-ACHE addition to the culture, ACHE mRNA levels dropped by approximately 90%, as compared with those in cells treated with the "sense" oligomer, S-ACHE. Four days after AS-ACHE treatment, ACHE mRNA increased to levels 10-fold higher than in S-ACHE cultures or in fresh bone marrow. At this later time point, differential PCR display revealed significant differences between cellular mRNA transcripts in bone marrow and those in AS-ACHE- or S-ACHE-treated cultures. These oligonucleotide-triggered effects underlay considerable alterations at the cellular level: AS-ACHE but not S-ACHE increased cell counts, reflecting enhanced proliferation. In the presence of erythropoietin it also enhanced colony counts, reflecting expansion of progenitors. AS-ACHE further suppressed apoptosis-related fragmentation of cellular DNA in the progeny cells, and it diverted hematopoiesis toward production of primitive blasts and macrophages in a dose-dependent manner promoted by erythropoietin. These findings suggest that the hematopoietic role of acetylcholinesterase, anticipated to be inverse to the observed antisense effects, is to reduce proliferation of the multipotent stem cells committed to erythropoiesis and megakaryocytopoiesis and macrophage production and to promote apoptosis in their progeny. Moreover, these findings may explain the tumorigenic association of perturbations in ACHE gene expression with leukemia.
ESTHER : Soreq_1994_Proc.Natl.Acad.Sci.U.S.A_91_7907
PubMedSearch : Soreq_1994_Proc.Natl.Acad.Sci.U.S.A_91_7907
PubMedID: 8058733

Title : Cholinotoxic effects on acetylcholinesterase gene expression are associated with brain-region specific alterations in G,C-rich transcripts - Lev-Lehman_1994_Brain.Res_661_75
Author(s) : Lev-Lehman E , el-Tamer A , Yaron A , Grifman M , Ginzberg D , Hanin I , Soreq H
Ref : Brain Research , 661 :75 , 1994
Abstract : To study the mechanisms underlying cholinotoxic brain damage, we examined ethylcholine aziridinium (AF64A) effects on cholinesterase genes. In vitro, AF64A hardly affected cholinesterase activities yet inhibited transcription of the G,C-rich AChE DNA encoding acetylcholinesterase (AChE) more than the A,T-rich butyrylcholinesterase (BChE) DNA. In vivo, intracerebroventricular injection of 2 nmol of AF64A decreased AChE mRNA in striatum and septum by 3- and 25-fold by day 7, with no change in BChE mRNA or AChE activity. In contrast, hippocampal AChE mRNA increased 10-fold by day 7 and BChE mRNA and AChE activity decreased 2-fold. By day 60 post-treatment, both AChE mRNA and AChE levels returned to normal in all regions except hippocampus, where AChE activity and BChE mRNA were decreased by 2-fold. Moreover, differential PCR displays revealed persistent induction, specific to the hippocampus of treated rats, of several unidentified G,C-rich transcripts, suggesting particular responsiveness of hippocampal G,C-rich genes to cholinotoxicity.
ESTHER : Lev-Lehman_1994_Brain.Res_661_75
PubMedSearch : Lev-Lehman_1994_Brain.Res_661_75
PubMedID: 7834388

Title : Testicular amplification and impaired transmission of human butyrylcholinesterase cDNA in transgenic mice - Beeri_1994_Hum.Reprod_9_284
Author(s) : Beeri R , Gnatt A , Lapidot-Lifson Y , Ginzberg D , Shani M , Soreq H , Zakut H
Ref : Hum Reprod , 9 :284 , 1994
Abstract : Gene amplification occurs frequently in tumour tissues yet is, in general, non-inheritable. To study the molecular mechanisms conferring this restraint, we created transgenic mice carrying a human butyrylcholinesterase (BCHE) coding sequence, previously found to be amplified in a father and son. Blot hybridization of tail DNA samples revealed somatic transgene amplifications with variable restriction patterns and intensities, suggesting the occurrence of independent amplification events, in 31% (11/35) of mice from the FII generation but in only 3.5% (2/58) of the FIII and FIV generations. In contrast, > 10-fold amplifications of the BCHE transgene and the endogenous acetylcholinesterase and c-raf genes appeared in both testis and epididymis DNA from > 80% of FIII mice. Drastic, selective reductions in testis BCHEmRNA but not in actin mRNA were detected by the PCR amplification of testis cDNA from the transgenic mice, and apparently resulted in the limited transmission of amplified genes. The testicular amplification of the BCHE transgene may potentially represent a general phenomenon with clinical implications in human infertility.
ESTHER : Beeri_1994_Hum.Reprod_9_284
PubMedSearch : Beeri_1994_Hum.Reprod_9_284
PubMedID: 8027285

Title : Expression of three alternative acetylcholinesterase messenger RNAs in human tumor cell lines of different tissue origins - Karpel_1994_Exp.Cell.Res_210_268
Author(s) : Karpel R , Ben-Aziz-Aloya R , Sternfeld M , Ehrlich G , Ginzberg D , Tarroni P , Clementi F , Zakut H , Soreq H
Ref : Experimental Cell Research , 210 :268 , 1994
Abstract : To study the molecular mechanisms underlying the intensive expression of acetylcholinesterase (AChE) in different tumor types, we characterized levels and composition of its messenger RNA (mRNA) sequences in heterologous tumor cell lines, primary tumor biopsies, and normal fetal and adult tissues and determined their exon-intron origin within the corresponding ACHE gene. Reverse transcription followed by polymerase chain reaction (RT-PCR) revealed three alternatively spliced ACHE mRNAs in NT2/D1 teratocarcinoma, NCI-N-592 small cell lung carcinoma, TE671 medulloblastoma, K-562 erythroleukemia, and 293 transformed embryonal kidney cells. The three ACHE mRNAs include the principal species expressed in brain and muscle and two additional transcripts containing insertions of 751 or 829 residues downstream from the exon 4 domain. The inserted region, which represents an intron in brain and muscle, is expressed in the tumor cell lines either as a "readthrough" form or with 78 residues deleted from its 5' end. A major band of 2.5 kb was labeled with ACHE cDNA in poly(A)+ RNA blots from medulloblastoma cells or brain tissue, whereas a PCR-amplified probe from the inserted domain labeled a 3.4-kb band but not the 2.5-kb band in poly(A)+ RNA from small cell lung carcinoma. The ACHE mRNAs including the alternative insertions were found only in cell lines with levels of the principal ACHE mRNA species equal to or higher than those in brain (1-10 molecules/cell), determined by following the kinetics of mRNA PCR amplification. Genomic DNA sequencing revealed that the inserted domains in the ACHE mRNAs expressed in the tumor cell lines encode C-terminal peptides of 40 and 14 residues. These include a free cysteine, terminate with the consensus HG element, and continue by a 29-residue-long C-terminal hydrophobic cleavable peptide, properties characteristic of precursors to phosphoinositide (PI)-linked proteins. In extension of the reported expression of PI-linked AChE in hemopoietic cells including K-562, our findings demonstrate the existence of ACHE mRNAs with the potential to encode one hydrophilic and two PI-linked forms of AChE in tumor cells from both hemopoietic and nonhemopoietic origins.
ESTHER : Karpel_1994_Exp.Cell.Res_210_268
PubMedSearch : Karpel_1994_Exp.Cell.Res_210_268
PubMedID: 8299725

Title : Population diversity and distinct haplotype frequencies associated with ACHE and BCHE genes of Israeli Jews from trans-Caucasian Georgia and from Europe - Ehrlich_1994_Genomics_22_288
Author(s) : Ehrlich G , Ginzberg D , Loewenstein Y , Glick D , Kerem B , Ben-Ari S , Zakut H , Soreq H
Ref : Genomics , 22 :288 , 1994
Abstract : Variant alleles of the butyrylcholinesterase gene, BCHE, have often been used to trace the genetic histories of populations. The D70G substitution in BCHE causes prolonged postanesthesia apnea ("atypical" phenotype); H322N substitution in the closely related acetylcholinesterase gene, ACHE, is the basis of the mutually incompatible Yt blood groups. In both genes, additional point mutations were reported to be linked to these phenotypically evident ones. To examine whether the intragenic linkage reported for the ACHE and BCHE mutations in Americans is universal, we studied frequencies of these mutations in trans-Caucasian Georgian Jews, a population that has remained relatively isolated for 1500 years. To this end we employed PCR amplification followed by DNA sequencing and enzymatic restriction and compared the frequencies we found to corresponding reported phenotype data. Georgian Jews' N322 ACHE was a rather low 7.0% and was totally linked to a P446 mutation, in agreement with a recent report. In BCHE, however, G70 was a relatively high 5.8%, and the V497 and T539 mutations were not found, either in Georgian or in Ashkenazi Jews, in contrast to reported findings in Americans. Our findings reveal distinct displays of ACHE and BCHE haplotypes in Georgian Jews and suggest different founder effects, genetic drifts, and/or selection pressures in the evolution of each of these genes.
ESTHER : Ehrlich_1994_Genomics_22_288
PubMedSearch : Ehrlich_1994_Genomics_22_288
PubMedID: 7806214

Title : Use of partially phosphorothioated antisense oligodeoxynucleotides for sequence-dependent modulation of hematopoiesis in culture - Ehrlich_1994_Antisense.Res.Dev_4_173
Author(s) : Ehrlich G , Patinkin D , Ginzberg D , Zakut H , Eckstein F , Soreq H
Ref : Antisense Research Development , 4 :173 , 1994
Abstract : To distinguish between sequence-dependent effects and non-specific cytotoxicity of phosphorothioate antisense oligonucleotides (AS-oligos), we introduced AS-oligos blocking expression of 2Hs, the Homo sapiens cell division controller cdc2 kinase, its hematopoietically expressed homolog CHED, and the acetylcholine-hydrolyzing enzyme butyrylcholinesterase (BCHE) into primary murine bone marrow (BM) culture. Antisense oligonucleotides were fully phosphorothioated (Ts) or prepared with three phosphorothioate groups at their 3' termini (S3). Each of these oligos could cause reductions in colony counts either as a result of its sequence-dependent biological capacity or due to sequence-independent cytotoxicity. The Ts and S3 forms of the matching sense oligo, S-BCHE, served for comparison. The S3 forms of AS-2Hs, AS-BCHE, and S-BCHE caused more limited drops in colony counts than their Ts counterparts, reflecting lower cytotoxicity. When incubated with electroblotted BM proteins, Ts but not S3 oligos intensively labeled two protein bands. Moreover, 5'-end 32P-labeled (Ts) S-BCHE labeled nuclear proteins in situ in small, mitotic cells, suggesting correlation between oligo-protein interactions and the sequence-independent cytotoxicity of Ts AS-oligos. Extension of the apparently nontoxic AS-CHED by two adenosine residues at the 3' end, creating a potential for intramolecular hydrogen bond formation, resulted in increased toxicity. These findings recommend the use of nonlooped, partially phosphorothioated oligos for the modulation of hematopoiesis.
ESTHER : Ehrlich_1994_Antisense.Res.Dev_4_173
PubMedSearch : Ehrlich_1994_Antisense.Res.Dev_4_173
PubMedID: 7849488

Title : Mapping the human acetylcholinesterase gene to chromosome 7q22 by fluorescent in situ hybridization coupled with selective PCR amplification from a somatic hybrid cell panel and chromosome-sorted DNA libraries - Ehrlich_1992_Genomics_13_1192
Author(s) : Ehrlich G , Viegas-Pequignot E , Ginzberg D , Sindel L , Soreq H , Zakut H
Ref : Genomics , 13 :1192 , 1992
Abstract : To establish the chromosomal location of the human ACHE gene encoding the acetylcholine hydrolyzing enzyme acetylcholinesterase (ACHE, acetylcholine acetylhydrolase, E.C., a human-specific polymerase chain reaction (PCR) procedure that supports the selective amplification of ACHE DNA fragments from human genomic DNA was employed with 19 human-hamster somatic cell hybrids carrying one or more human chromosomes. Informative ACHE-specific PCR fragments were produced from two cell lines, both of which include human chromosome 7, but not with DNA from 17 cell hybrids carrying various combinations of all human chromosomes other than 7. Fluorescent in situ hybridization of biotinylated ACHE DNA with metaphase chromosomes from human peripheral blood lymphocytes revealed prominent labeling on the 7q22 position. Therefore, further tests were performed to confirm the chromosome 7 location. DNA samples from the two cell lines including chromosome 7 and the ACHE gene were positive with PCR primers informative for the human cystic fibrosis CFTR gene, known to reside at the 7q31.1 position, but negative for the ACHE-related butyrylcholinesterase (BCHE, acylcholine acylhydrolase, E.C. gene, mapped at the 3q26-ter position, confirming that these lines contain chromosome 7 but not chromosome 3. In contrast, three other cell lines including chromosome 3, but not 7, were BCHE-positive and ACHE-negative. In addition, genomic DNA from a sorted chromosome 7 library supported the production of ACHE- but not BCHE-specific PCR products, whereas with DNA from a sorted chromosome 3 library, the BCHE but not the ACHE fragment was amplified.
ESTHER : Ehrlich_1992_Genomics_13_1192
PubMedSearch : Ehrlich_1992_Genomics_13_1192
PubMedID: 1380483

Title : Chorionic villus cDNA library displays expression of butyrylcholinesterase: putative genetic disposition for ecological danger - Zakut_1991_Prenat.Diagnos_11_597
Author(s) : Zakut H , Lieman-Hurwitz J , Zamir R , Sindell L , Ginzberg D , Soreq H
Ref : Prenat Diagnos , 11 :597 , 1991
Abstract : Gene expression in chorionic villi may be particularly vulnerable to environmental exposure to poisonous substances. To reveal villus gene products which are thus subject to poisoning, molecular cloning was employed. A single sample of apparently normal chorionic villi (approximately 40 mg, from 9 weeks' gestation) was microscopically dissected to ensure purity of fetal tissue. Total RNA was extracted by isothiocyanate and directly employed for reverse transcription. A chorionic villus cDNA library was constructed from this preparation in the phage vector lambda gt10 and contained 60,000 independent recombinants. In the present study, this cDNA library was screened with labelled cDNA probes encoding human butyrylcholinesterase (BCHE) and acetylcholinesterase (ACHE). Nine BCHEcDNA clones were isolated out of 1.6 x 10(6) phages (5.7 x 10(-6) of screened recombinants) and exhibited similar restriction patterns to those observed for BCHEcDNA from other human tissues. In contrast, no ACHEcDNA clones could be found in 4.0 x 10(6) screened phages (less than 2.5 x 10(-6) of recombinants). These findings demonstrate efficient transcription (similar to fetal brain levels) from the BCHE gene but not from the ACHE gene in chorionic villi, and support the notion that BCHE is involved in chorionic villus growth and development.
ESTHER : Zakut_1991_Prenat.Diagnos_11_597
PubMedSearch : Zakut_1991_Prenat.Diagnos_11_597
PubMedID: 1722578

Title : Differential Codon Usage and Distinct Surface Probabilities in Human Acetylcholinesterase and Butyrylcholinesterase -
Author(s) : Ben-Aziz R , Gnatt A , Prody C , Lev-Lehman E , Neville LF , Seidman S , Ginzberg D , Soreq H , Lapidot-Lifson Y , Zakut H
Ref : In: Cholinesterases: Structure, Function, Mechanism, Genetics, and Cell Biology , (Massoulie J, Barnard EA, Chatonnet A, Bacou F, Doctor BP, Quinn DM) American Chemical Society, Washington, DC :172 , 1991

Title : Human acetylcholinesterase and butyrylcholinesterase are encoded by two distinct genes - Gnatt_1991_Cell.Mol.Neurobiol_11_91
Author(s) : Gnatt A , Ginzberg D , Lieman-Hurwitz J , Zamir R , Zakut H , Soreq H
Ref : Cellular Molecular Neurobiology , 11 :91 , 1991
Abstract : 1. Various hybridization approaches were employed to investigate structural and chromosomal interrelationships between the human cholinesterase genes CHE and ACHE encoding the polymorphic, closely related, and coordinately regulated enzymes having butyrylcholinesterase (BCHE) and acetylcholinesterase (AChE) activities. 2. Homologous cosmid recombination with a 190-base pair 5' fragment from BCHEcDNA resulted in the isolation of four overlapping cosmid clones, apparently derived from a single gene with several introns. The Cosmid CHEDNA included a 700-base pair fragment known to be expressed at the 3' end of BCHEcDNA from nervous system tumors and which has been mapped by in situ hybridization to the unique 3q26-ter position. In contrast, cosmid CHEDNA did not hybridize with full-length AChEcDNA, proving that the complete CHE gene does not include AChE-encoding sequences either in exons or in its introns. 3. The chromosomal origin of BCHE-encoding sequences was further examined by two unrelated gene mapping approaches. Filter hybridization with DNA from human/hamster hybrid cell lines revealed BCHEcDNA-hybridizing sequences only in cell lines including human chromosome 3. However, three BCHEcDNA-homologous sequences were observed at chromosomal positions 3q21, 3q26-ter, and 16q21 by a highly stringent in situ hybridization protocol, including washes at high temperature and low salt. 4. These findings stress the selectivity of cosmid recombination and chromosome blots, raise the possibility of individual differences in BCHEcDNA-hybridizing sequences, and present an example for a family highly similar proteins encoded by distinct, nonhomologous genes.
ESTHER : Gnatt_1991_Cell.Mol.Neurobiol_11_91
PubMedSearch : Gnatt_1991_Cell.Mol.Neurobiol_11_91
PubMedID: 2013062

Title : Acetylcholinesterase and butyrylcholinesterase genes coamplify in primary ovarian carcinomas - Zakut_1990_J.Clin.Invest_86_900
Author(s) : Zakut H , Ehrlich G , Ayalon A , Prody CA , Malinger G , Seidman S , Ginzberg D , Kehlenbach R , Soreq H
Ref : Journal of Clinical Investigation , 86 :900 , 1990
Abstract : The genes for acetylcholinesterase (ACHE) and butyrylcholinesterase (CHE) are expressed in multiple tumor tissues, including ovarian carcinomas. Both CHE and ACHE genes coamplify in leukemias. To examine the relationship of gene amplification to the expression of these genes in tumors, ACHE and CHE genes and their expression were studied in primary ovarian carcinomas. DNA blot hybridization demonstrated a significant amplification and mutagenesis of both genes in 6 of 11 malignant tumors studied. This was greater or of the same order of magnitude as the amplification of the oncogenes c-rafi, v-sis, and c-fes in these tumors. No amplification was found in normal ovarian tissues or benign ovarian cysts. Xenopus oocyte microinjections, blot and in situ hybridizations, and immuno- and cytochemical staining revealed translatable CHEmRNA and its active protein product in discrete tumor foci. The frequent coamplification in ovarian carcinomas of ACHE and CHE genes implicates cholinesterases in neoplastic growth and/or proliferation.
ESTHER : Zakut_1990_J.Clin.Invest_86_900
PubMedSearch : Zakut_1990_J.Clin.Invest_86_900
PubMedID: 2394839

Title : Molecular cloning and construction of the coding region for human acetylcholinesterase reveals a G + C-rich attenuating structure - Soreq_1990_Proc.Natl.Acad.Sci.U.S.A_87_9688
Author(s) : Soreq H , Ben-Aziz R , Prody CA , Seidman S , Gnatt A , Neville LF , Lieman-Hurwitz J , Lev-Lehman E , Ginzberg D , Lipidot-Lifson Y
Ref : Proceedings of the National Academy of Sciences of the United States of America , 87 :9688 , 1990
Abstract : To study the primary structure of human acetylcholinesterase (AcChoEase; EC and its gene expression and amplification, cDNA libraries from human tissues expressing oocyte-translatable AcChoEase mRNA were constructed and screened with labeled oligodeoxynucleotide probes. Several cDNA clones were isolated that encoded a polypeptide with greater than or equal to 50% identically aligned amino acids to Torpedo AcChoEase and human butyrylcholinesterase (BtChoEase; EC However, these cDNA clones were all truncated within a 300-nucleotide-long G + C-rich region with a predicted pattern of secondary structure having a high Gibbs free energy (-117 kcal/mol) downstream from the expected 5' end of the coding region. Screening of a genomic DNA library revealed the missing 5' domain. When ligated to the cDNA and constructed into a transcription vector, this sequence encoded a synthetic mRNA translated in microinjected oocytes into catalytically active AcChoEase with marked preference for acetylthiocholine over butyrylthiocholine as a substrate, susceptibility to inhibition by the AcChoEase inhibitor BW284C51, and resistance to the BtChoEase inhibitor tetraisopropylpyrophosphoramide. Blot hybridization of genomic DNA from different individuals carrying amplified AcChoEase genes revealed variable intensities and restriction patterns with probes from the regions upstream and downstream from the predicted G + C-rich structure. Thus, the human AcChoEase gene includes a putative G + C-rich attenuator domain and is subject to structural alterations in cases of AcChoEase gene amplification.
ESTHER : Soreq_1990_Proc.Natl.Acad.Sci.U.S.A_87_9688
PubMedSearch : Soreq_1990_Proc.Natl.Acad.Sci.U.S.A_87_9688
PubMedID: 2263619
Gene_locus related to this paper: human-ACHE

Title : Coamplification of human acetylcholinesterase and butyrylcholinesterase genes in blood cells: correlation with various leukemias and abnormal megakaryocytopoiesis - Lapidot-Lifson_1989_Proc.Natl.Acad.Sci.U.S.A_86_4715
Author(s) : Lapidot-Lifson Y , Prody CA , Ginzberg D , Meytes D , Zakut H , Soreq H
Ref : Proceedings of the National Academy of Sciences of the United States of America , 86 :4715 , 1989
Abstract : To study the yet unknown role of the ubiquitous family of cholinesterases (ChoEases) in developing blood cells, the recently isolated cDNAs encoding human acetylcholinesterase (AcChoEase; acetylcholine acetylhydrolase, EC and butyrylcholinesterase (BtChoEase; cholinesterase; acylcholine acylhydrolase, EC were used in blot hybridization with peripheral blood DNA from various leukemic patients. Hybridization signals (10- to 200-fold intensified) and modified restriction patterns were observed with both cDNA probes in 4 of the 16 leukemia DNA preparations examined. These reflected the amplification of the corresponding AcChoEase and BtChoEase genes (ACHE and CHE) and alteration in their structure. Parallel analysis of 30 control samples revealed nonpolymorphic, much weaker hybridization signals for each of the probes. In view of previous reports on the effect of acetylcholine analogs and ChoEase inhibitors in the induction of megakaryocytopoiesis and production of platelets in the mouse, we further searched for such phenomena in nonleukemic patients with platelet production disorders. Amplifications of both ACHE and CHE genes were found in 2 of the 4 patients so far examined. Pronounced coamplification of these two related but distinct genes in correlation with pathological production of blood cells suggests a functional role for members of the ChoEase family in megakaryocytopoiesis and raises the question whether the coamplification of these genes could be causally involved in the etiology of hemocytopoietic disorders.
ESTHER : Lapidot-Lifson_1989_Proc.Natl.Acad.Sci.U.S.A_86_4715
PubMedSearch : Lapidot-Lifson_1989_Proc.Natl.Acad.Sci.U.S.A_86_4715
PubMedID: 2734315