[Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]

K. L. Brodolin; V. M. Studitskii; A. D. Mirzabekov

[Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]

K. L. Brodolin, V. M. Studitskii, A. D. Mirzabekov

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

Abstract

The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown.

Original language	Russian
Pages (from-to)	1085-1093
Number of pages	9
Journal	Molekulyarnaya Biologiya
Volume	27
Issue number	5
State	Published - 1993

Keywords

Cross-Linking Reagents DNA-Binding Proteins/*chemistry DNA-Directed RNA Polymerases/*chemistry Escherichia coli/*enzymology Formaldehyde/*chemistry *Promoter Regions, Genetic

Cite this

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title = "[Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]",

abstract = "The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown.",

keywords = "Cross-Linking Reagents DNA-Binding Proteins/*chemistry DNA-Directed RNA Polymerases/*chemistry Escherichia coli/*enzymology Formaldehyde/*chemistry *Promoter Regions, Genetic",

author = "Brodolin, {K. L.} and Studitskii, {V. M.} and Mirzabekov, {A. D.}",

note = "Brodolin, K L Studitskii, V M Mirzabekov, A D Journal Article Russia (Federation) 1993/09/01 Mol Biol (Mosk). 1993 Sep-Oct;27(5):1085-93.",

year = "1993",

language = "Russian",

volume = "27",

pages = "1085--1093",

journal = "Molekulyarnaya Biologiya",

issn = "0026-8984",

publisher = "Russian Academy of Sciences",

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TY - JOUR

T1 - [Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]

AU - Brodolin, K. L.

AU - Studitskii, V. M.

AU - Mirzabekov, A. D.

N1 - Brodolin, K L Studitskii, V M Mirzabekov, A D Journal Article Russia (Federation) 1993/09/01 Mol Biol (Mosk). 1993 Sep-Oct;27(5):1085-93.

PY - 1993

Y1 - 1993

N2 - The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown.

AB - The protein-protein and DNA-protein crosslinks produced by formaldehyde were used to investigate the intersubunit and subunit-DNA interactions for free RNA polymerase and for an open complex of RNA polymerase with the lacUV5 promoter. In both cases the contacts between beta,beta' and beta', sigma subunits were observed, while there were no contacts between beta and sigma subunits. Only one of beta or beta' subunits and a sigma subunit crosslink to promoter DNA. We have chosen the conditions for fixing the RNA polymerase-DNA complexes on different stages of transcription initiation. The possibility to use limited fixation with low concentrations of formaldehyde to study specific DNA-protein interactions was shown.

KW - Cross-Linking Reagents DNA-Binding Proteins/chemistry DNA-Directed RNA Polymerases/chemistry Escherichia coli/enzymology Formaldehyde/chemistry Promoter Regions, Genetic

M3 - Article

SN - 0026-8984

VL - 27

SP - 1085

EP - 1093

JO - Molekulyarnaya Biologiya

JF - Molekulyarnaya Biologiya

IS - 5

ER -

[Study of the structure of Escherichia coli RNA polymerase and its complex with the lacUV5-promotor using protein-protein and DNA-protein crosslinks, formed by formaldehyde]

Abstract

Keywords

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