Study of DNA topoisomerase II in ataxia-telangiectasia cells.
We have provided evidence recently for a defect in DNA topoisomerase II in ataxia--telangiectasia (A-T) lymphoblastoid cells. This study was initiated to investigate in greater detail the nature of this defect. Southern hybridization analysis was carried out on DNA from control and A-T Epstein--Barr virus-transformed lymphoblastoid cells. The pattern of digestion, using several restriction enzymes, was the same in both cell types. Expression of topoisomerase II mRNA occurred to the same extent and there was no difference in the size of mRNA between the cell types. Western blot analysis revealed that the same amount of a major band of topoisomerase II protein was present in A-T and control cells but there was evidence for a reduced amount of a lower-molecular-weight form in A-T only. Extraction and purification did not lead to alteration in size of the enzyme or in amount recovered.
Singh
S P
SP
Queensland Institute of Medical Research, Australia.
Lavin
M F
MF
D001260:Ataxia Telangiectasia / Q000201:enzymology*; D015152:Blotting, Northern; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics; D012016:Reference Values; D014158:Transcription, Genetic
D012333:RNA, Messenger; D012313:RNA; D004247:DNA; D004250:DNA Topoisomerases, Type II