Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex
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Uranyl mediated photofootprinting reveals strong E. coli RNA polymerase--DNA backbone contacts in the +10 region of the DeoP1 promoter open complex. / Jeppesen, C; Nielsen, Peter E.
I: Nucleic Acids Research, Bind 17, Nr. 13, 11.07.1989, s. 4947-56.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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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 - Jeppesen, C
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
U2 - 10.1093/nar/17.13.4947
DO - 10.1093/nar/17.13.4947
M3 - Journal article
C2 - 2503811
VL - 17
SP - 4947
EP - 4956
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 13
ER -
ID: 203632221