The VirD2 Protein of Agrobacterium Tumefaciens Carries Nuclear Localization Signals Important for Transfer of T-DNA to Plant

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1993 Jun 1

PMID 8391110

Citations 46

Authors

L Rossi

B Hohn

B Tinland

Affiliations

Soon will be listed here.

Abstract

Agrobacterium tumefaciens is able to transfer a piece of DNA, the T-DNA, to the nucleus of the plant cell. The VirD2 protein is required for the production of the T-DNA, it is tightly linked to the T-DNA and it is thought to direct it to the plant genome. Two nuclear localization signals (NLS), one in the N-terminal part and one in the C-terminal part of the VirD2 protein, have been shown to be able to target marker proteins to the plant nucleus. Here we analyze nuclear entry of the T-DNA complex using a new and very sensitive assay for T-DNA transfer. We show that optimal T-DNA transfer requires the VirD2 NLS located in the C-terminal part of the protein, whereas mutations in the N-terminal NLS coding sequence seem to have no effect on T-DNA transfer.

Citing Articles

Genomic consequences associated with Agrobacterium-mediated transformation of plants.

Thomson G, Dickinson L, Jacob Y Plant J. 2023; 117(2):342-363.

PMID: 37831618 PMC: 10841553. DOI: 10.1111/tpj.16496.

Modeling emergence of toxin-antidote protein functions with an evolutionary algorithm.

Beckmann J, Gillespie J, Tauritz D Front Microbiol. 2023; 14:1116766.

PMID: 37362913 PMC: 10288140. DOI: 10.3389/fmicb.2023.1116766.

Modelling Emergence of Toxin-Antidote Protein Functions with an Evolutionary Algorithm.

Beckmann J, Gillespie J, Tauritz D bioRxiv. 2023; .

PMID: 36993585 PMC: 10055314. DOI: 10.1101/2023.03.23.533954.

JAZ8 Interacts With VirE3 Attenuating Agrobacterium Mediated Root Tumorigenesis.

Li S, Xu B, Niu X, Lu X, Cheng J, Zhou M Front Plant Sci. 2021; 12:685533.

PMID: 34868098 PMC: 8639510. DOI: 10.3389/fpls.2021.685533.

Increased Agrobacterium-mediated transformation of Saccharomyces cerevisiae after deletion of the yeast ADA2 gene.

Roushan M, Shao S, Poledri I, Hooykaas P, van Heusden G Lett Appl Microbiol. 2021; 74(2):228-237.

PMID: 34816457 PMC: 9299121. DOI: 10.1111/lam.13605.

References

van der Krol A, Chua N . The basic domain of plant B-ZIP proteins facilitates import of a reporter protein into plant nuclei. Plant Cell. 1991; 3(7):667-75. PMC: 160034. DOI: 10.1105/tpc.3.7.667. View

Hooykaas P, Schilperoort R . Agrobacterium and plant genetic engineering. Plant Mol Biol. 1992; 19(1):15-38. DOI: 10.1007/BF00015604. View

Mozo T, Hooykaas P . Factors affecting the rate of T-DNA transfer from Agrobacterium tumefaciens to Nicotiana glauca plant cells. Plant Mol Biol. 1992; 19(6):1019-30. DOI: 10.1007/BF00040533. View

Raineri D, Stephens K, Ramos C, Tinland B, Nester E, Hohn B . 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. J Bacteriol. 1993; 175(3):723-31. PMC: 196211. DOI: 10.1128/jb.175.3.723-731.1993. View

Steck T, Morel P, Kado C . Vir box sequences in Agrobacterium tumefaciens pTiC58 and A6. Nucleic Acids Res. 1988; 16(17):8736. PMC: 338614. DOI: 10.1093/nar/16.17.8736. View

Bonnard G, Tinland B, Paulus F, Szegedi E, Otten L . Nucleotide sequence, evolutionary origin and biological role of a rearranged cytokinin gene isolated from a wide host range biotype III Agrobacterium strain. Mol Gen Genet. 1989; 216(2-3):428-38. DOI: 10.1007/BF00334387. View

Steck T, Lin T, Kado C . VirD2 gene product from the nopaline plasmid pTiC58 has at least two activities required for virulence. Nucleic Acids Res. 1990; 18(23):6953-8. PMC: 332755. DOI: 10.1093/nar/18.23.6953. View

Parke D . Construction of mobilizable vectors derived from plasmids RP4, pUC18 and pUC19. Gene. 1990; 93(1):135-7. DOI: 10.1016/0378-1119(90)90147-j. View

Tinland B, Hall M, Hohn B . The T-DNA-linked VirD2 protein contains two distinct functional nuclear localization signals. Proc Natl Acad Sci U S A. 1992; 89(16):7442-6. PMC: 49726. DOI: 10.1073/pnas.89.16.7442. View

10.

Henkel T, Zabel U, Van Zee K, Muller J, Fanning E, Baeuerle P . Intramolecular masking of the nuclear location signal and dimerization domain in the precursor for the p50 NF-kappa B subunit. Cell. 1992; 68(6):1121-33. DOI: 10.1016/0092-8674(92)90083-o. View

11.

Dingwall C, Laskey R . Nuclear targeting sequences--a consensus?. Trends Biochem Sci. 1991; 16(12):478-81. DOI: 10.1016/0968-0004(91)90184-w. View

12.

Stachel S, Nester E . The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J. 1986; 5(7):1445-54. PMC: 1166964. DOI: 10.1002/j.1460-2075.1986.tb04381.x. View

13.

Puchta H, Hohn B . A transient assay in plant cells reveals a positive correlation between extrachromosomal recombination rates and length of homologous overlap. Nucleic Acids Res. 1991; 19(10):2693-700. PMC: 328188. DOI: 10.1093/nar/19.10.2693. View

14.

Rihs H, Jans D, Fan H, Peters R . The rate of nuclear cytoplasmic protein transport is determined by the casein kinase II site flanking the nuclear localization sequence of the SV40 T-antigen. EMBO J. 1991; 10(3):633-9. PMC: 452694. DOI: 10.1002/j.1460-2075.1991.tb07991.x. View

15.

Citovsky V, Wong M, Zambryski P . Cooperative interaction of Agrobacterium VirE2 protein with single-stranded DNA: implications for the T-DNA transfer process. Proc Natl Acad Sci U S A. 1989; 86(4):1193-7. PMC: 286652. DOI: 10.1073/pnas.86.4.1193. View

16.

Steck T, Close T, Kado C . High levels of double-stranded transferred DNA (T-DNA) processing from an intact nopaline Ti plasmid. Proc Natl Acad Sci U S A. 1989; 86(7):2133-7. PMC: 286865. DOI: 10.1073/pnas.86.7.2133. View

17.

Kalderon D, Richardson W, Markham A, Smith A . Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984; 311(5981):33-8. DOI: 10.1038/311033a0. View

18.

Robbins J, Dilworth S, Laskey R, Dingwall C . Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell. 1991; 64(3):615-23. DOI: 10.1016/0092-8674(91)90245-t. View

19.

Mattanovich D, Ruker F, Machado A, Laimer M, Regner F, Steinkellner H . Efficient transformation of Agrobacterium spp. by electroporation. Nucleic Acids Res. 1989; 17(16):6747. PMC: 318385. DOI: 10.1093/nar/17.16.6747. View

20.

Shurvinton C, Hodges L, Ream W . A nuclear localization signal and the C-terminal omega sequence in the Agrobacterium tumefaciens VirD2 endonuclease are important for tumor formation. Proc Natl Acad Sci U S A. 1992; 89(24):11837-41. PMC: 50652. DOI: 10.1073/pnas.89.24.11837. View