Document Detail


Oligomerization of the E. coli core RNA polymerase: formation of (α2ββ'ω)2-DNA complexes and regulation of the oligomerization by auxiliary subunits.
MedLine Citation:
PMID:  21533049     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In this work, using multiple, dissimilar physico-chemical techniques, we demonstrate that the Escherichia coli RNA polymerase core enzyme obtained through a classic purification procedure forms stable (α(2)ββ'ω)(2) complexes in the presence or absence of short DNA probes. Multiple control experiments indicate that this self-association is unlikely to be mediated by RNA polymerase-associated non-protein molecules. We show that the formation of (α(2)ββ'ω)(2) complexes is subject to regulation by known RNA polymerase interactors, such as the auxiliary SWI/SNF subunit of RNA polymerase RapA, as well as NusA and σ(70). We also demonstrate that the separation of the core RNA polymerase and RNA polymerase holoenzyme species during Mono Q chromatography is likely due to oligomerization of the core enzyme. We have analyzed the oligomeric state of the polymerase in the presence or absence of DNA, an aspect that was missing from previous studies. Importantly, our work demonstrates that RNA polymerase oligomerization is compatible with DNA binding. Through in vitro transcription and in vivo experiments (utilizing a RapA(R599/Q602) mutant lacking transcription-stimulatory function), we demonstrate that the formation of tandem (α(2)ββ'ω)(2)-DNA complexes is likely functionally significant and beneficial for the transcriptional activity of the polymerase. Taken together, our findings suggest a novel structural aspect of the E. coli elongation complex. We hypothesize that transcription by tandem RNA polymerase complexes initiated at hypothetical bidirectional "origins of transcription" may explain recurring switches of the direction of transcription in bacterial genomes.
Authors:
Seema G Kansara; Maxim V Sukhodolets
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2011-04-20
Journal Detail:
Title:  PloS one     Volume:  6     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2011  
Date Detail:
Created Date:  2011-05-02     Completed Date:  2011-08-30     Revised Date:  2014-09-21    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e18990     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Base Sequence
Chromatography, Gel
DNA Probes
DNA, Bacterial / chemistry,  metabolism*
DNA-Directed RNA Polymerases / chemistry,  metabolism*
Escherichia coli / enzymology*
Molecular Sequence Data
Molecular Weight
Nucleic Acid Conformation
Grant Support
ID/Acronym/Agency:
R15 GM081803/GM/NIGMS NIH HHS; R15 GM081803-01/GM/NIGMS NIH HHS; R15GM081803/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/DNA Probes; 0/DNA, Bacterial; EC 2.7.7.6/DNA-Directed RNA Polymerases
Comments/Corrections

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