Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex

Peter E. Nielsen

Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex

Transcription, RNA, and Gene Medicine Program

38 Citationer (Scopus)

Abstract

Employing a newly developed uranyl photofootprinting technique (Nielsen et al. (1988) FEBS Lett. 235, 122), we have analyzed the structure of the E. coli RNA polymerase deoP1 promoter open complex. The results show strong polymerase DNA backbone contacts in the -40, -10, and most notably in the +10 region. These results suggest that unwinding of the -12 to +3 region of the promoter in the open complex is mediated through polymerase DNA backbone contacts on both sides of this region. The pattern of bases that are hyperreactive towards KMnO4 or uranyl within the -12 to +3 region furthermore argues against a model in which this region is simply unwound and/or single stranded. The results indicate specific protein contacts and/or a fixed DNA conformation within the -12 to +3 region.

Originalsprog	Engelsk
Tidsskrift	Nucleic Acids Research
Vol/bind	17
Udgave nummer	13
Sider (fra-til)	4947-56
Antal sider	10
ISSN	0305-1048
Status	Udgivet - 11 jul. 1989

Citationsformater

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title = "Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex",

abstract = "Employing a newly developed uranyl photofootprinting technique (Nielsen et al. (1988) FEBS Lett. 235, 122), we have analyzed the structure of the E. coli RNA polymerase deoP1 promoter open complex. The results show strong polymerase DNA backbone contacts in the -40, -10, and most notably in the +10 region. These results suggest that unwinding of the -12 to +3 region of the promoter in the open complex is mediated through polymerase DNA backbone contacts on both sides of this region. The pattern of bases that are hyperreactive towards KMnO4 or uranyl within the -12 to +3 region furthermore argues against a model in which this region is simply unwound and/or single stranded. The results indicate specific protein contacts and/or a fixed DNA conformation within the -12 to +3 region.",

keywords = "Base Sequence, DNA, Bacterial/genetics, DNA-Directed RNA Polymerases/metabolism, Edetic Acid, Escherichia coli/enzymology, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Nucleotide Mapping/methods, Plasmids, Promoter Regions, Genetic, Protein Binding, Uranium",

author = "Nielsen, {Peter E.}",

year = "1989",

month = jul,

day = "11",

language = "English",

volume = "17",

pages = "4947--56",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "13",

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TY - JOUR

T1 - Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex

AU - Nielsen, Peter E.

PY - 1989/7/11

Y1 - 1989/7/11

N2 - Employing a newly developed uranyl photofootprinting technique (Nielsen et al. (1988) FEBS Lett. 235, 122), we have analyzed the structure of the E. coli RNA polymerase deoP1 promoter open complex. The results show strong polymerase DNA backbone contacts in the -40, -10, and most notably in the +10 region. These results suggest that unwinding of the -12 to +3 region of the promoter in the open complex is mediated through polymerase DNA backbone contacts on both sides of this region. The pattern of bases that are hyperreactive towards KMnO4 or uranyl within the -12 to +3 region furthermore argues against a model in which this region is simply unwound and/or single stranded. The results indicate specific protein contacts and/or a fixed DNA conformation within the -12 to +3 region.

AB - Employing a newly developed uranyl photofootprinting technique (Nielsen et al. (1988) FEBS Lett. 235, 122), we have analyzed the structure of the E. coli RNA polymerase deoP1 promoter open complex. The results show strong polymerase DNA backbone contacts in the -40, -10, and most notably in the +10 region. These results suggest that unwinding of the -12 to +3 region of the promoter in the open complex is mediated through polymerase DNA backbone contacts on both sides of this region. The pattern of bases that are hyperreactive towards KMnO4 or uranyl within the -12 to +3 region furthermore argues against a model in which this region is simply unwound and/or single stranded. The results indicate specific protein contacts and/or a fixed DNA conformation within the -12 to +3 region.

KW - Base Sequence

KW - DNA, Bacterial/genetics

KW - DNA-Directed RNA Polymerases/metabolism

KW - Edetic Acid

KW - Escherichia coli/enzymology

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Nucleic Acid Conformation

KW - Nucleotide Mapping/methods

KW - Plasmids

KW - Promoter Regions, Genetic

KW - Protein Binding

KW - Uranium

M3 - Journal article

C2 - 2503811

SN - 0305-1048

VL - 17

SP - 4947

EP - 4956

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 13

ER -

Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex

Abstract

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