Proteogenomics in microbiology: Taking the right turn at the junction of genomics and proteomics

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Abstract
High‐accuracy and high‐throughput proteomic methods have completely changed the way we can identify and characterize proteins. MS‐based proteomics can now provide a unique supplement to genomic data and add a new level of information to the interpretation of genomic sequences. Proteomics‐driven genome annotation has become especially relevant in microbiology where genomes are sequenced on a daily basis and limitations of an in silico driven annotation process are well recognized. In this review paper, we outline different strategies on how one can design a proteogenomic experiment, for example on genome‐sequenced (synonymous proteogenomics) versus unsequenced organisms (ortho‐proteogenomics) or with the aid of other “omic” data such as RNA‐seq. We touch upon many challenges that are encountered during a typical proteogenomic study, mostly concerning bioinformatics methods and downstream data analysis, but also related to creation and use of sequence databases. A large list of proteogenomic case studies of different microorganisms is provided to illustrate the mapping of MS/MS‐derived peptide spectra to genomic DNA sequences. These investigations have led to accurate determination of translational initiation sites, pointed out eventual read‐throughs or programmed frameshifts, detected signal peptide processing or other protein maturation events, removed questionable annotation assignments, and provided evidence for predicted hypothetical proteins.
Funding Information
  • Research Council of Norway (204743).)

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