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PMID: 1579485 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

KorB protein of promiscuous plasmid RP4 recognizes inverted sequence repetitions in regions essential for conjugative plasmid transfer.

Nucleic acids research ·Vol. 20 ·No. 8 ·1992-04-25 ·Pages 1851-8

Balzer D, Ziegelin G, Pansegrau W, Kruft V, Lanka E

Abstract

We have constructed a RP4 KorB overproducing strain and purified the protein to near homogeneity. KorB is a DNA binding protein recognizing defined palindromic 13-bp sequences (TTTAGCSGCTAAA). Inverted sequence repetitions of this type, designated OB, are present on RP4 12 times. OB-sequences are localized in replication and maintenance regions as well as in the regions Tra1 and Tra2 essential for conjugative transfer. All sites found in Tra regions by computer search act as targets for specific binding of KorB protein. KorB-DNA complexes were detected by DNA fragment retardation assay using polyacrylamide gels. The 13-bp symmetric arrangement of the consensus OB-sequence constitutes the core for binding KorB protein since any truncation of this sequence prevents complex assembly or leads to a considerable destabilization of the KorB-DNA complexes. A hydroxyl radical footprint analysis demonstrated complex formation of KorB with the OB-sequence directly and suggests the presence of an unusual DNA structure within the nucleoprotein complex.

Related Genes
korB
MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,metabolism Base Sequence Binding Sites/genetics Cloning, Molecular Conjugation, Genetic Consensus Sequence/genetics DNA/metabolism Escherichia coli/genetics Molecular Sequence Data Nucleic Acid Conformation Plasmids/genetics Repetitive Sequences, Nucleic Acid/genetics Repressor Proteins/genetics,metabolism
Chemicals
Bacterial Proteins KorB protein, Plasmid RK2 Repressor Proteins DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Balzer D
Max-Planck-Institut für Molekulare Genetik, Berlin, Germany.
Ziegelin G
Pansegrau W
Kruft V
Lanka E
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/20.8.1851
Published
1992-04-25
Pages
1851-8
Language
English
Region
England
NLM ID
0411011
PMCID
PMC312297
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/20.8.1851 PMC Full Text View journal details
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