The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease

Yanofsky, Martin F.; Porter, Stephen C.; Young, C C; Albright, Lisa M.; Gordon, Milton P.; Nester, Eugene W.

doi:10.1016/0092-8674(86)90604-5

Cited by 244 publications

(214 citation statements)

References 34 publications

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“…Bacterial (LB) and yeast (YNB supplemented with the required amino acids) media were prepared as described (28,30). Plasmid pMY1153 contains the virD region ofthe octopine Ti plasmid pTiA6NC (31). pTTO is a pUC derivative containing the promoter and the terminator for the 35S RNA of cauliflower mosaic virus (CaMV) (32).…”

Section: Methodsmentioning

confidence: 99%

The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals.

Tinland

Koukolíková-Nicola

Hall

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

130

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Agrobacterium tumefaciens causes neoplastic growth in plants by transferring a piece ofDNA, called T-DNA, into the nucleus of the plant cell. The virulence protein VirD2 ofA. tumefaciens is tightly linked to the T-DNA and is thought to direct it to the plant genome. Here we show that the VirD2 protein contains two nuclear localization signals that are functional both in yeast and in plant cells. One signal is located in the N-terminal part of the protein and resembles a singlecluster-type nuclear localization signal. The second signal is near the C terminus and is a bipartite-type nuclear localization signal. The involvement of these sequences in the entry of the T-DNA into the nucleus is discussed.Agrobacterium tumefaciens, the causative agent of crown gall disease, transfers into plant cells genes coding for enzymes involved in the synthesis of plant growth factors. These genes are carried by the T-DNA, a well-defined region of a large plasmid called Ti (tumor-inducing). The T-DNA is delimited at both extremities by two almost perfect 25-basepair repeats, or border sequences (for reviews see refs.

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Section: Methodsmentioning

confidence: 99%

The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals.

Tinland

Koukolíková-Nicola

Hall

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

130

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“…In vivo in Agrobacterium, VirD1/D2 are both required for the nicking reaction (Scheiffele et al, 1995;Stachel et al, 1987;Yanofsky et al, 1986). In vitro, VirD2 alone nicks ss oligonucleotides bearing the T-DNA border sequence (Jasper et al, 1994;Pansegrau et al, 1993a).…”

Section: Nic Sites and Nicking Enzymes (Table 1 Item 8)mentioning

confidence: 99%

The transfer of DNA from Agrobacterium tumefaciens into plants: a feast of fundamental insights

et al. 2000

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This review is dedicated to Jeff Schell, one of the founders of modern`Agrobiology', the genetic and molecular dissection of crown gall disease. Together with notable scientists at the University of Gent, Belgium, Jeff spearheaded the discovery of the Ti-plasmid. The elusive`tumor inducing principle' was uncloaked and provided impetus for an incredibly fruitful subsequent 25 years of analyses. Scientists all over the world were caught up in unraveling the underlying mechanisms of Agrobacterium-mediated gene transfer to plants, and along the way uncovered a movable feast of fundamental insights. Below we summarize a sampling of Agrobacterium's most recently recognized accomplishments.

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“…For A. tumefaciens T-DNA transfer system, this subset comprises the VirD2 relaxase, VirD1 and VirC proteins, and may also include VirE2, VirE1, and VirF (4,5,7). VirD2 cleaves the bottom strand of the T-DNA at the two border sequences to generate singlestranded (ss) T-strand (8,9) and remains covalently associated with the 5Ј end of the T-strand (10). VirE2 is a nonspecific ssDNAbinding protein that can coat the length of the T-strand in vitro (11).…”

mentioning

confidence: 99%

Recruitment of conjugative DNA transfer substrate to Agrobacterium type IV secretion apparatus

Guo

Jin

Sun

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Bacterial type IV secretion system (T4SS) belongs to a growing class of evolutionarily conserved transporters that translocate DNA and proteins into a wide variety of organisms including bacterial and eukaryotic cells. Archetypal is the Agrobacterium tumefaciens VirB/D4 T4SS that transfers oncogenic T-DNA to various eukaryotic cells, which is transferred as a nucleoprotein T-complex with VirD2 as the pilot protein. As a derivative of plasmid conjugation systems, the VirB/D4 T4SS can also transfer certain mobilizable plasmids and bacterial proteins like VirE2 and VirF, although it is unknown how the membrane-bound T4SS recruits different transfer substrates. Here, we show that a cytoplasmic VirD2-binding protein (VBP) is involved in the recruitment of the T-complex to the energizing components of the T4SS, including VirD4, VirB4, and VirB11. VBP is also important for the recruitment of a conjugative plasmid to a different transfer system independent of VirB/D4. These data indicate that VBP functions as a previously unrecognized recruiting protein that helps couple nucleoprotein substrates to the appropriate transport sites for conjugative DNA transfers. VBP has three functionally redundant homologs, and similar homologs can be found in different bacterial genomes, suggesting a previously uncharacterized class of proteins involved in conjugative DNA transfers.bacterial type IV secretion system ͉ T-DNA ͉ virulence

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The virD operon of Agrobacterium tumefaciens encodes a site-specific endonuclease

Cited by 244 publications

References 34 publications

The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals.

The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals.

The transfer of DNA from Agrobacterium tumefaciens into plants: a feast of fundamental insights

Recruitment of conjugative DNA transfer substrate to Agrobacterium type IV secretion apparatus

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