Gallien_2009_Genome.Res_19_128

The progress in sequencing technologies irrigates biology with an ever-increasing number of genome sequences In most cases the gene repertoire is predicted in silico and conceptually translated into proteins As recently highlighted the predicted genes exhibit frequent errors particularly in start codons with a serious impact on subsequent biological studies A new ortho-proteogenomic approach is presented here for the annotation refinement of multiple genomes at once It combines comparative genomics with an original proteomic protocol that allows the characterization of both N-terminal and internal peptides in a single experiment This strategy was applied to the Mycobacterium genus with Mycobacterium smegmatis as the reference and identified 946 distinct proteins including 443 characterized N termini These experimental data allowed the correction of 19 of the characterized start codons the identification of 29 proteins missed during the annotation process and the curation thanks to comparative genomics of 4328 sequences of 16 other Mycobacterium proteomes.