Molecular characterization of DNA repair protein Ku70 from Vitis vinifera and its purification from transgenic tobacco.
The DNA double strand break repair in plants is preferentially by non homologous end joining (NHEJ) pathway. A key protein of NHEJ pathway is Ku70. We have identified Ku70 homolog (VvKu70) from grapevine genome database. In this report we characterize a Ku70 homologue from Vitis vinifera cv. Mango. The VvKu70 expression was found to increase strongly in response to gamma radiation. The transcript level of VvKu70 was found to increase up to 36 h in gamma irradiated shoots of grapevine. The expression of VvKu70 was found in many organs like stem, leaves and roots. A GFP fused VvKu70 protein was found to be nuclear localized which indicates that the VvKu70 is a nuclear localized protein. The VvKu70 identified by in silico approaches is present as a single copy number in V. vinifera cv. Mango genome. The VvKu70-GFP fused protein possesses ATPase activity and fails to bind dsDNA but binds ssDNA.
Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India. hsjtak@barc.gov.in
Himanshu
H
Tak
Minal
M
Mhatre
D000818:Animals; D034961:Antigens, Nuclear / Q000235:genetics / Q000302:isolation & purification*; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects; D053904:DNA Breaks, Single-Stranded / Q000528:radiation effects; D059766:DNA End-Joining Repair / Q000528:radiation effects; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification*; D005720:Gamma Rays; D018506:Gene Expression Regulation, Plant / Q000528:radiation effects; D000072200:Ku Autoantigen; D018515:Plant Leaves / Q000235:genetics / Q000528:radiation effects; D018517:Plant Roots / Q000235:genetics / Q000528:radiation effects; D030821:Plants, Genetically Modified / Q000528:radiation effects*; D014026:Nicotiana / Q000235:genetics*; D027843:Vitis / Q000235:genetics / Q000528:radiation effects
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D000072200:Ku Autoantigen