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PMID: 2836366 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The Agrobacterium tumefaciens virE2 gene product is a single-stranded-DNA-binding protein that associates with T-DNA.

Journal of bacteriology ·Vol. 170 ·No. 6 ·1988-06-00 ·Pages 2659-67

Christie PJ, Ward JE, Winans SC, Nester EW

Abstract

Agrobacterium tumefaciens transfers T-DNA into the plant genome by a process mediated by Ti plasmid-encoded vir genes. Cleavage at T-DNA border sequences by the VirD endonuclease generates linear, single-stranded T-DNA molecules. In the work described in this report, we used electrophoretic mobility shift assays to show that the purified virE2 gene product binds to single-stranded DNA. VirE2 protein associates with T-DNA as shown by immunoprecipitation studies with VirE2-specific antiserum. The VirE2 protein was detected primarily in the cytoplasm, but also in the inner and outer membrane and periplasmic fractions. Virulence of a virE2 mutant was restored by mixed infection with strains carrying an intact vir region, but not with virA, virB, virD, virE, or virG mutants or chvA, chvB, or exoC mutants. We propose that the VirE2 protein is involved in the processing of T-DNA and in T-strand protection during transfer to the plant cell.

MeSH Terms
Bacterial Proteins/analysis,metabolism Base Sequence DNA, Bacterial/metabolism DNA-Binding Proteins/analysis,metabolism Electrophoresis, Polyacrylamide Gel Escherichia coli/analysis,genetics Plasmids Rhizobium/genetics,pathogenicity
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins T-DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Christie P J
Department of Microbiology, University of Washington, Seattle 98195.
Ward J E
Winans S C
Nester E W
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
Published
1988-06-00
Pages
2659-67
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC211185
Subset
IM
Grants
NIGMS NIH HHS · 5 RO1 GM 32618-14 · United States
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