Conformational changes in E.coli RNA polymerase during promoter recognition

Konstantin L. Brodolin; Vasily M. Studitsky; Andrei D. Mirzabekov

doi:10.1093/nar/21.24.5748

Conformational changes in E.coli RNA polymerase during promoter recognition

Konstantin L. Brodolin, Vasily M. Studitsky, Andrei D. Mirzabekov

RAS - Engelhardt Institute of Molecular Biology

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

We analysed complexes formed during recognition of the lacUV5 promoter by E.coli RNA polymerase using formaldehyde as a DNA - protein and protein - protein cross-linking reagent. Most of the cross-linked complexes specific for the open complex (RP_O) contain the β′ subunit of RNA polymerase cross-linked with promoter DNA in the regions: - 50 to - 49; - 5 to -10; +5 to +8 and +18 to +21. The protein - protein cross-linking pattern of contacting subunits is the same for the RNA polymerase in solution and in RP0: there are strong α-β′ and β-β′ interactions. In contrast, only β-β′ cross-links were detected in the closed (RP_c) and intermediate (RP_l) complexes. In presence of lac repressor before or after formation of the RP_o crosslinking pattern is similar with that of RP_l (RP_c) complex.

Original language	English
Pages (from-to)	5748-5753
Number of pages	6
Journal	Nucleic Acids Research
Volume	21
Issue number	24
DOIs	https://doi.org/10.1093/nar/21.24.5748
State	Published - Dec 11 1993

Keywords

DNA-Directed RNA Polymerases/chemistry
Electrophoresis, Polyacrylamide Gel
Escherichia coli/enzymology
Promoter Regions, Genetic
Protein Conformation
Repressor Proteins/metabolism
Solutions
Temperature

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title = "Conformational changes in E.coli RNA polymerase during promoter recognition",

abstract = "We analysed complexes formed during recognition of the lacUV5 promoter by E.coli RNA polymerase using formaldehyde as a DNA - protein and protein - protein cross-linking reagent. Most of the cross-linked complexes specific for the open complex (RPO) contain the β′ subunit of RNA polymerase cross-linked with promoter DNA in the regions: - 50 to - 49; - 5 to -10; +5 to +8 and +18 to +21. The protein - protein cross-linking pattern of contacting subunits is the same for the RNA polymerase in solution and in RP0: there are strong α-β′ and β-β′ interactions. In contrast, only β-β′ cross-links were detected in the closed (RPc) and intermediate (RPl) complexes. In presence of lac repressor before or after formation of the RPo crosslinking pattern is similar with that of RPl (RPc) complex.",

keywords = "DNA-Directed RNA Polymerases/chemistry, Electrophoresis, Polyacrylamide Gel, Escherichia coli/enzymology, Promoter Regions, Genetic, Protein Conformation, Repressor Proteins/metabolism, Solutions, Temperature",

author = "Brodolin, {Konstantin L.} and Studitsky, {Vasily M.} and Mirzabekov, {Andrei D.}",

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AU - Brodolin, Konstantin L.

AU - Studitsky, Vasily M.

AU - Mirzabekov, Andrei D.

PY - 1993/12/11

Y1 - 1993/12/11

N2 - We analysed complexes formed during recognition of the lacUV5 promoter by E.coli RNA polymerase using formaldehyde as a DNA - protein and protein - protein cross-linking reagent. Most of the cross-linked complexes specific for the open complex (RPO) contain the β′ subunit of RNA polymerase cross-linked with promoter DNA in the regions: - 50 to - 49; - 5 to -10; +5 to +8 and +18 to +21. The protein - protein cross-linking pattern of contacting subunits is the same for the RNA polymerase in solution and in RP0: there are strong α-β′ and β-β′ interactions. In contrast, only β-β′ cross-links were detected in the closed (RPc) and intermediate (RPl) complexes. In presence of lac repressor before or after formation of the RPo crosslinking pattern is similar with that of RPl (RPc) complex.

AB - We analysed complexes formed during recognition of the lacUV5 promoter by E.coli RNA polymerase using formaldehyde as a DNA - protein and protein - protein cross-linking reagent. Most of the cross-linked complexes specific for the open complex (RPO) contain the β′ subunit of RNA polymerase cross-linked with promoter DNA in the regions: - 50 to - 49; - 5 to -10; +5 to +8 and +18 to +21. The protein - protein cross-linking pattern of contacting subunits is the same for the RNA polymerase in solution and in RP0: there are strong α-β′ and β-β′ interactions. In contrast, only β-β′ cross-links were detected in the closed (RPc) and intermediate (RPl) complexes. In presence of lac repressor before or after formation of the RPo crosslinking pattern is similar with that of RPl (RPc) complex.

KW - DNA-Directed RNA Polymerases/chemistry

KW - Electrophoresis, Polyacrylamide Gel

KW - Escherichia coli/enzymology

KW - Promoter Regions, Genetic

KW - Protein Conformation

KW - Repressor Proteins/metabolism

KW - Solutions

KW - Temperature

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JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 24

ER -

Conformational changes in E.coli RNA polymerase during promoter recognition

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