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PMID: 7505669 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

NusG alters rho-dependent termination of transcription in vitro independent of kinetic coupling.

Gene expression ·Vol. 3 ·No. 2 ·1993-00-00 ·Pages 119-33

Nehrke KW, Zalatan F, Platt T

Abstract

To complement the recent discovery that rho-dependent termination in E. coli requires nusG protein in vivo, we have tested the effect of purified nusG protein on rho-dependent termination in vitro. With the well-characterized trp t' terminator of E. coli, and no other proteins than E. coli RNA polymerase and rho factor, nusG causes a proximal shift in the terminated RNA endpoints, compared to the endpoints generated by rho alone. The presence of nusG also enhances rho-mediated termination on partially defective mutant trp t' templates. We rule out explanations such as a change in the kinetic coupling between rho and RNA polymerase or a nusG-mediated increase in the affinity of rho for RNA. We also detect no difference in the helicase rate of rho in the presence of nusG. Even assays with completely stalled and isolated ternary complexes indicate that rho is able to effect the release of RNA with the assistance of nusG at points preceding the most proximal release sites observed in the absence of nusG. Our observations support a model in which nusG acts as a component of the transcription complex, possibly interacting with both rho and RNA polymerase as it governs accessibility to the nascent transcript.

MeSH Terms
Bacterial Proteins/metabolism Base Sequence DNA, Bacterial DNA-Directed RNA Polymerases/metabolism Escherichia coli/genetics,metabolism Escherichia coli Proteins Kinetics Molecular Sequence Data Peptide Elongation Factors/metabolism Point Mutation RNA, Bacterial/metabolism Rho Factor/metabolism Suppression, Genetic Terminator Regions, Genetic Transcription Factors Transcription, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial Escherichia coli Proteins NusG protein, E coli Peptide Elongation Factors RNA, Bacterial Rho Factor Transcription Factors DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Nehrke K W
Department of Biochemistry, University of Rochester Medical Center, NY 14642.
Zalatan F
Platt T
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44 references, click to expand
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Article Info
Journal
Gene expression
Abbr.
Gene Expr
ISSN
1052-2166
Published
1993-00-00
Pages
119-33
Language
English
Region
United States
NLM ID
9200651
PMCID
PMC6081637
Subset
IM
Grants
NIAID NIH HHS · T32 AI007362 · United States
NIAID NIH HHS · 1T32AI-07362 · United States
NIGMS NIH HHS · 5-RO1-GM35658 · United States
External Links
PubMed source PMC Full Text View journal details
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