Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.
An assay has been developed in which excision repair deficiency of xeroderma pigmentosum cells is transiently complemented, as measured by unscheduled DNA synthesis, by microinjection of cytoplasmic poly(A)+ RNA derived from HeLa cells. Four different complementation groups of xeroderma pigmentosum have been assayed. Groups A and G showed complementation, whereas groups D and F did not. Survival for cells in each of the groups subsequent to microinjection was approximately equal to 75%. Approximately 10-25% of surviving cells from groups A and G were complemented, as judged by near-normal unscheduled DNA synthesis. Fractionation of cytoplasmic poly(A)+ RNA on a 15-30% nondenaturing sucrose gradient and subsequent microinjection of the individual fractions indicate that repair mRNAs that complement xeroderma pigmentosum groups A and G sediment at approximately 11 S and 12 S, respectively. This assay should be of great utility in the cloning and biochemical analysis of DNA repair genes.
D001345:Autoradiography; D002460:Cell Line; D004261:DNA Replication; D005816:Genetic Complementation Test; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D008845:Microinjections; D011061:Poly A / Q000235:genetics* / Q000302:isolation & purification / Q000528:radiation effects; D012313:RNA / Q000235:genetics* / Q000302:isolation & purification / Q000528:radiation effects; D012333:RNA, Messenger; D014316:Tritium; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D012333:RNA, Messenger; D014316:Tritium; D011061:Poly A; D012313:RNA