RRC ID 35527
Author Chen SS, Sperling E, Silverman JM, Davis JH, Williamson JR.
Title Measuring the dynamics of E. coli ribosome biogenesis using pulse-labeling and quantitative mass spectrometry.
Journal Mol Biosyst
Abstract The ribosome is an essential organelle responsible for cellular protein synthesis. Until recently, the study of ribosome assembly has been largely limited to in vitro assays, with few attempts to reconcile these results with the more complex ribosome biogenesis process inside the living cell. Here, we characterize the ribosome synthesis and assembly pathway for each of the E. coli ribosomal protein (r-protein) in vivo using a stable isotope pulse-labeling timecourse. Isotope incorporation into assembled ribosomes was measured by quantitative mass spectrometry (qMS) and fit using steady-state flux models. Most r-proteins exhibit precursor pools ranging in size from 0% to 7% of completed ribosomes, and the sizes of these individual r-protein pools correlate well with the order of r-protein binding in vitro. Additionally, we observe anomalously large precursor pools for specific r-proteins with known extra-ribosomal functions, as well as three r-proteins that apparently turnover during steady-state growth. Taken together, this highly precise, time-dependent proteomic qMS approach should prove useful in future studies of ribosome biogenesis and could be easily extended to explore other complex biological processes in a cellular context.
Volume 8(12)
Pages 3325-34
Published 2012-10-30
DOI 10.1039/c2mb25310k
PMID 23090316
PMC PMC3501348
MeSH Escherichia coli / genetics Escherichia coli / metabolism* Escherichia coli Proteins / analysis Escherichia coli Proteins / genetics Escherichia coli Proteins / metabolism* Mass Spectrometry Nitrogen Isotopes Proteomics RNA, Ribosomal / chemistry RNA, Ribosomal / genetics RNA, Ribosomal / metabolism Ribosomal Proteins / chemistry Ribosomal Proteins / metabolism* Ribosome Subunits, Large, Bacterial / metabolism* Ribosome Subunits, Small, Bacterial / metabolism* Ribosomes / metabolism*
IF 2.855
Times Cited 24
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Prokaryotes E. coli ?