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PMID: 1917866 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specific DNA binding of the TraM protein to the oriT region of plasmid R100.

Journal of bacteriology ·Vol. 173 ·No. 20 ·1991-10-00 ·Pages 6347-54

Abo T, Inamoto S, Ohtsubo E

Abstract

The product of the traM gene of plasmid R100 was purified as the TraM-collagen-beta-galactosidase fusion protein (TraM*) by using a beta-galactosidase-specific affinity column, and the TraM portion of TraM* (TraM') was separated by collagenolysis. Both the TraM* and TraM' proteins were found to bind specifically to a broad region preceding the traM gene. This region (designated sbm) was located within the nonconserved region in oriT among conjugative plasmids related to R100. The region seems to contain four core binding sites (designated sbmA, sbmB, sbmC, and sbmD), each consisting of a similar number of nucleotides and including a homologous 15-bp sequence. This result, together with the observation that the TraM* protein was located in the membrane fraction, indicates the possibility that the TraM protein has a function in anchoring the oriT region of R100 at the sbm sites to the membrane pore, through which the single-stranded DNA is transferred to the recipient. sbmC and sbmD, each of which contained a characteristic inverted repeat sequence, overlapped with the promoter region for the traM gene. This suggests that the expression of the traM gene may be regulated by its own product.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,isolation & purification,metabolism Base Sequence Binding Sites/genetics Chromatography, Affinity Collagen/genetics,metabolism Conjugation, Genetic DNA Replication/genetics DNA, Bacterial/metabolism DNA, Circular/metabolism DNA-Binding Proteins/genetics,isolation & purification,metabolism Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics,metabolism Molecular Sequence Data R Factors Recombinant Fusion Proteins/biosynthesis Sequence Alignment beta-Galactosidase/genetics
Chemicals
Bacterial Proteins DNA, Bacterial DNA, Circular DNA-Binding Proteins Recombinant Fusion Proteins TraM protein, bacterial Collagen beta-Galactosidase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Abo T
Institute of Applied Microbiology, University of Tokyo, Japan.
Inamoto S
Ohtsubo E
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30 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.20.6347-6354.1991
Published
1991-10-00
Pages
6347-54
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208966
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.173.20.6347-6354.1991 PMC Full Text View journal details
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