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PMID: 23542154 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Dynamics of translation by single ribosomes through mRNA secondary structures.

Nature structural & molecular biology ·Vol. 20 ·No. 5 ·2013-05-00 ·Pages 582-8

Chen C, Zhang H, Broitman SL, Reiche M, Farrell I, Cooperman BS, Goldman YE

Abstract

During protein synthesis, the ribosome translates nucleotide triplets in single-stranded mRNA into polypeptide sequences. Strong downstream mRNA secondary structures, which must be unfolded for translation, can slow or even halt protein synthesis. Here we used single-molecule fluorescence resonance energy transfer to determine reaction rates for specific steps within the elongation cycle as the Escherichia coli ribosome encounters stem-loop or pseudoknot mRNA secondary structures. Downstream stem-loops containing 100% GC base pairs decrease the rates of both tRNA translocation within the ribosome and deacylated tRNA dissociation from the ribosomal exit site (E site). Downstream stem-loops or pseudoknots containing both GC and AU pairs also decrease the rate of tRNA dissociation, but they have little effect on tRNA translocation rate. Thus, somewhat unexpectedly, unfolding of mRNA secondary structures is more closely coupled to E-site tRNA dissociation than to tRNA translocation.

MeSH Terms
Escherichia coli/genetics,metabolism Fluorescence Resonance Energy Transfer Kinetics Models, Biological Models, Molecular Nucleic Acid Conformation Protein Biosynthesis RNA, Messenger/metabolism Ribosomes/metabolism
Chemicals
RNA, Messenger
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Chen Chunlai
Pennsylvania Muscle Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Zhang Haibo
Broitman Steven L
Reiche Michael
Farrell Ian
Cooperman Barry S
Goldman Yale E
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Article Info
Journal
Nature structural & molecular biology
Abbr.
Nat Struct Mol Biol
ISSN
1545-9985
DOI
10.1038/nsmb.2544
Published
2013-05-00
Epub
2013-00-31
Pages
582-8
Language
English
Region
United States
NLM ID
101186374
PMCID
PMC3648610
Subset
IM
Grants
NIGMS NIH HHS · R01 GM080376 · United States
NIGMS NIH HHS · R01GM080376 · United States
External Links
PubMed source DOI: 10.1038/nsmb.2544 PMC Full Text View journal details
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