Non-homologous end-joining proteins are required for Agrobacterium T-DNA integration.
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.
van Attikum
H
H
Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands.
Bundock
P
P
Hooykaas
P J
PJ
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D034961:Antigens, Nuclear; D001483:Base Sequence; D002869:Chromosome Aberrations; D004265:DNA Helicases; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000502:physiology; D004269:DNA, Bacterial / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000502:physiology; D005090:Exodeoxyribonucleases / Q000502:physiology; D005656:Fungal Proteins / Q000502:physiology; D005822:Genetic Vectors; D005838:Genotype; D000072200:Ku Autoantigen; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000502:physiology; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae; D016615:Telomere / Q000378:metabolism
D034961:Antigens, Nuclear; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C000607891:DNL4 protein, S cerevisiae; D005656:Fungal Proteins; C000607893:LIG4 protein, human; D009687:Nuclear Proteins; C086930:RAD50 protein, S cerevisiae; C061573:SIR4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae; C036483:T-DNA; C086929:XRS2 protein, S cerevisiae; C520769:YKU70 protein, S cerevisiae; C081647:high affinity DNA-binding factor, S cerevisiae; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; C094191:MRE11 protein, S cerevisiae; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen; D011088:DNA Ligases; D000072481:DNA Ligase ATP