Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration.
The transferred DNA (T-DNA) is transported from Agrobacterium tumefaciens to the nucleus and is stably integrated into the genome of many plant species. It has been proposed that the VirD2 protein, tightly attached to the T-DNA, pilots the T-DNA into the plant cell nucleus and that it is involved in integration. Using agroinfection and beta-glucuronidase expression as two different very sensitive transient assays for T-DNA transfer, together with assays for stable integration, we have shown that the C-terminal half of the VirD2 protein and the VirD3 protein are not involved in T-DNA integration. However, the bipartite nuclear localization signal, which is located within the C terminus of the VirD2 protein and which has previously been shown to be able to target a foreign protein into the plant cell nucleus, was shown to be required for efficient T-DNA transfer. virD4 mutants were shown by agroinfection to be completely inactive in T-DNA transfer.
Friedrich Miescher-Institut, Basel, Switzerland.
Z
Z
Koukolíková-Nicola
D
D
Raineri
K
K
Stephens
C
C
Ramos
B
B
Tinland
E W
EW
Nester
B
B
Hohn
D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000472:pathogenicity; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009876:Operon; D010944:Plants / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic; D017384:Sequence Deletion
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides