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PMID: 14633991 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Transcript cleavage factors GreA and GreB act as transient catalytic components of RNA polymerase.

The EMBO journal ·Vol. 22 ·No. 23 ·2003-12-01 ·Pages 6322-34

Laptenko O, Lee J, Lomakin I, Borukhov S

Abstract

Prokaryotic transcription elongation factors GreA and GreB stimulate intrinsic nucleolytic activity of RNA polymerase (RNAP). The proposed biological role of Gre-induced RNA hydrolysis includes transcription proofreading, suppression of transcriptional pausing and arrest, and facilitation of RNAP transition from transcription initiation to transcription elongation. Using an array of biochemical and molecular genetic methods, we mapped the interaction interface between Gre and RNAP and identified the key residues in Gre responsible for induction of nucleolytic activity in RNAP. We propose a structural model in which the C-terminal globular domain of Gre binds near the opening of the RNAP secondary channel, the N-terminal coiled-coil domain (NTD) protrudes inside the RNAP channel, and the tip of the NTD is brought to the immediate vicinity of RNAP catalytic center. Two conserved acidic residues D41 and E44 located at the tip of the NTD assist RNAP by coordinating the Mg2+ ion and water molecule required for catalysis of RNA hydrolysis. If so, Gre would be the first transcription factor known to directly participate in the catalytic act of RNAP.

MeSH Terms
Amino Acid Sequence Catalysis Cross-Linking Reagents DNA-Directed RNA Polymerases/chemistry,genetics Endoribonucleases/genetics Escherichia coli/genetics,metabolism Escherichia coli Proteins/chemistry,genetics Hydroxyl Radical/metabolism Models, Molecular Molecular Sequence Data Protein Conformation Protein Subunits Sequence Alignment Sequence Homology, Amino Acid Transcription Factors/chemistry,genetics Transcription, Genetic Transcriptional Elongation Factors
Chemicals
Cross-Linking Reagents Escherichia coli Proteins GreA protein, E coli GreB protein, E coli Protein Subunits Transcription Factors Transcriptional Elongation Factors Hydroxyl Radical DNA-Directed RNA Polymerases Endoribonucleases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Laptenko Oleg
Department of Microbiology and Immunology, SUNY Health Science Center at Brooklyn, 450 Clarkson Avenue, BSB 3-27, Brooklyn, NY 11203, USA.
Lee Jookyung
Lomakin Ivan
Borukhov Sergei
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg610
Published
2003-12-01
Pages
6322-34
Language
English
Region
England
NLM ID
8208664
PMCID
PMC291851
Subset
IM
Grants
NIGMS NIH HHS · R01 GM054098 · United States
NIGMS NIH HHS · GM54098 · United States
External Links
PubMed source DOI: 10.1093/emboj/cdg610 PMC Full Text View journal details
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