Mode of DNA binding by SopA protein of Escherichia coli F plasmid.

Abstract:

:The binding of SopA to the promoter region of its own gene, in which four copies of SopA's recognition sequence, 5'-CTTTGC-3', are arrayed asymmetrically, was examined in vitro. Titration using electrophoretic mobility shift assay showed that the stoichiometry of SopA protomers to the promoter-region DNA is 4 and that the binding is highly co-operative. The co-operativity was corroborated by EMSA and DNase I footprinting for a number of mutant DNA fragments in which 5'-CTTTGC-3' was changed to 5'-CTTACG-3'. EMSA in the style of circular permutation showed that SopA bends DNA. Mutation at either outermost binding site had a different effect on DNA bending by SopA, reflecting the asymmetry in the arrangement of the binding sites, for which the results of DNase I footprinting were in agreement. Gel filtration chromatography and analytical ultracentrifugation of free SopA showed that the protein can exist as a monomer and oligomers in the absence of ATP. Hence, the results indicate that the co-operativity in SopA's DNA binding is based on its intrinsic protein-protein interaction modified by DNA interaction.

journal_name

J Biochem

journal_title

Journal of biochemistry

authors

Komai M,Umino M,Hanai R

doi

10.1093/jb/mvq151

subject

Has Abstract

pub_date

2011-04-01 00:00:00

pages

455-61

issue

4

eissn

0021-924X

issn

1756-2651

pii

mvq151

journal_volume

149

pub_type

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