With the rapid increase in the number of sequenced prokaryotic genomes, relying on automated gene annotation became a necessity. Multiple lines of evidence, however, suggest that current bacterial genome annotations may contain inconsistencies and are incomplete, even for so-called well-annotated genomes. We here discuss underexplored sources of protein diversity and new methodologies for high-throughput genome reannotation. The expression of multiple molecular forms of proteins (proteoforms) from a single gene, particularly driven by alternative translation initiation, is gaining interest as a prominent contributor to bacterial protein diversity. In consequence, riboproteogenomic pipelines were proposed to comprehensively capture proteoform expression in prokaryotes by the complementary use of (positional) proteomics and the direct readout of translated genomic regions using ribosome profiling. To complement these discoveries, tailored strategies are required for the functional characterization of newly discovered bacterial proteoforms.

中文翻译：

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细菌核糖体基因组学：揭示了N末端蛋白形式存在的时代。

随着测序的原核基因组数目的迅速增加，依靠自动基因注释成为必要。但是，有多种证据表明，即使对于所谓的注释正确的基因组，当前的细菌基因组注释也可能包含不一致且不完整。我们在这里讨论蛋白质多样性的探索不足的来源和高通量基因组重注释的新方法。作为单个细菌蛋白质多样性的重要贡献者，单个基因的多种分子形式的蛋白质（蛋白形式）的表达，尤其是由交替翻译启动驱动，已引起人们的关注。结果，提出了核糖基因组学管道，以通过互补使用（位置）蛋白质组学和使用核糖体谱分析直接读出翻译的基因组区域来全面捕获原核生物中的蛋白形式表达。为了补充这些发现，需要针对新发现的细菌蛋白形式的功能表征采用量身定制的策略。