Clarifying the Translational Pausing Landscape in Bacteria by Ribosome Profiling

Fuad Mohammad, Christopher Woolstenhulme, Rachel Green, Allen Buskirk

Research output: Contribution to journalArticle

Abstract

The rate of protein synthesis varies according to the mRNA sequence in ways that affect gene expression. Global analysis of translational pausing is now possible with ribosome profiling. Here, we revisit an earlier report that Shine-Dalgarno sequences are the major determinant of translational pausing in bacteria. Using refinements in the profiling method as well as biochemical assays, we find that SD motifs have little (if any) effect on elongation rates. We argue that earlier evidence of pausing arose from two factors. First, in previous analyses, pauses at Gly codons were difficult to distinguish from pauses at SD motifs. Second, and more importantly, the initial study preferentially isolated long ribosome-protected mRNA fragments that are enriched in SD motifs. These findings clarify the landscape of translational pausing in bacteria as observed by ribosome profiling.

Original languageEnglish (US)
Pages (from-to)686-694
Number of pages9
JournalCell Reports
Volume14
Issue number4
DOIs
StatePublished - Feb 2 2016

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Ribosomes
Bacteria
Messenger RNA
Gene expression
Elongation
Assays
Codon
Gene Expression
Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Clarifying the Translational Pausing Landscape in Bacteria by Ribosome Profiling. / Mohammad, Fuad; Woolstenhulme, Christopher; Green, Rachel; Buskirk, Allen.

In: Cell Reports, Vol. 14, No. 4, 02.02.2016, p. 686-694.

Research output: Contribution to journalArticle

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