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PMID: 11707425 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Non-homologous end-joining proteins are required for Agrobacterium T-DNA integration.

The EMBO journal ·Vol. 20 ·No. 22 ·2001-11-15 ·Pages 6550-8

van Attikum H, Bundock P, Hooykaas PJ

Abstract

Agrobacterium tumefaciens causes crown gall disease in dicotyledonous plants by introducing a segment of DNA (T-DNA), derived from its tumour-inducing (Ti) plasmid, into plant cells at infection sites. Besides these natural hosts, Agrobacterium can deliver the T-DNA also to monocotyledonous plants, yeasts and fungi. The T-DNA integrates randomly into one of the chromosomes of the eukaryotic host by an unknown process. Here, we have used the yeast Saccharomyces cerevisiae as a T-DNA recipient to demonstrate that the non-homologous end-joining (NHEJ) proteins Yku70, Rad50, Mre11, Xrs2, Lig4 and Sir4 are required for the integration of T-DNA into the host genome. We discovered a minor pathway for T-DNA integration at the telomeric regions, which is still operational in the absence of Rad50, Mre11 or Xrs2, but not in the absence of Yku70. T-DNA integration at the telomeric regions in the rad50, mre11 and xrs2 mutants was accompanied by gross chromosomal rearrangements.

MeSH Terms
Agrobacterium tumefaciens/genetics Antigens, Nuclear Base Sequence Chromosome Aberrations DNA Helicases DNA Ligase ATP DNA Ligases/physiology DNA, Bacterial/genetics,metabolism DNA-Binding Proteins/physiology DNA-Directed DNA Polymerase/metabolism Electrophoresis, Polyacrylamide Gel Endodeoxyribonucleases/physiology Exodeoxyribonucleases/physiology Fungal Proteins/physiology Genetic Vectors Genotype Ku Autoantigen Models, Genetic Molecular Sequence Data Mutation Nuclear Proteins/physiology Polymerase Chain Reaction Protein Binding Recombination, Genetic Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins Silent Information Regulator Proteins, Saccharomyces cerevisiae Telomere/metabolism
Chemicals
Antigens, Nuclear DNA, Bacterial DNA-Binding Proteins DNL4 protein, S cerevisiae Fungal Proteins LIG4 protein, human Nuclear Proteins RAD50 protein, S cerevisiae SIR4 protein, S cerevisiae Saccharomyces cerevisiae Proteins Silent Information Regulator Proteins, Saccharomyces cerevisiae T-DNA XRS2 protein, S cerevisiae YKU70 protein, S cerevisiae high affinity DNA-binding factor, S cerevisiae bacteriophage T7 induced DNA polymerase DNA-Directed DNA Polymerase Endodeoxyribonucleases Exodeoxyribonucleases MRE11 protein, S cerevisiae DNA Helicases XRCC5 protein, human Xrcc6 protein, human Ku Autoantigen DNA Ligases DNA Ligase ATP
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
van Attikum H
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands.
Bundock P
Hooykaas P J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.22.6550
Published
2001-11-15
Pages
6550-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125718
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.22.6550 PMC Full Text View journal details
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