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Label-free shotgun proteomics of tmRNA substrates enriched by immunoprecipitation

ABSTRACT: We used a combination of genetic and proteomic approaches to characterize tmRNA (ssrA) activity in the genome-reduced bacterium Mycoplasma pneumoniae. For this, we generated tmRNA mutants encoding a tag resistant to proteolysis. Endogenous protein tagging by the mutant tmRNA gene (ssrAmk) was then examined by immunoprecipitation (IP) enrichment followed by LC-MS/MS analysis.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mycoplasma Pneumoniae (strain Atcc 29342 / M129)

SUBMITTER: Marc Weber

LAB HEAD: Luis Serrano

PROVIDER: PXD023973 | Pride | 2021-08-23

REPOSITORIES: Pride

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Widespread ribosome stalling in a genome-reduced bacterium and the need for translational quality control.

Burgos Raul R Weber Marc M Gallo Carolina C Lluch-Senar Maria M Serrano Luis L

iScience 20210816 9

Trans-translation is a ubiquitous bacterial mechanism of ribosome rescue mediated by a transfer-messenger RNA (tmRNA) that adds a degradation tag to the truncated nascent polypeptide. Here, we characterize this quality control system in a genome-reduced bacterium, <i>Mycoplasma pneumoniae</i> (MPN), and perform a comparative analysis of protein quality control components in slow and fast-growing prokaryotes. We show <i>in vivo</i> that in MPN the sole quality control cytoplasmic protease (Lon) d ...[more]

PMID: 34485867

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