Pulse-labeled, small closed circular DNA in cultured mouse and human cells.
Mouse (erythroleukemia, TSA8, and FM3A) cells and human (HeLa and HL-60) cells were pulse-labeled with [3H]thymidine and covalently closed circular DNA in the extrachromosomal fraction was analyzed by fluorography following polyacrylamide gel electrophoresis. Two discrete bands for mouse and at least one, different, band for human cells emerged in the position to which small circular DNA (less than 1 kb) migrate, suggesting there to be species-specific, preferentially labeled, small circular DNA in mammalian cells. The incorporation of [3H]thymidine into the DNA was inhibited by cycloheximide but unaffected by aphidicolin. Restriction enzyme (AluI) digestion of the DNA fraction from MEL cells produced approximately 120-, 100-, and 50-bp labeled DNA fragments. The origin of the pulse-labeled DNAs is discussed.
Kiyama
R
R
Institute of Applied Microbiology, University of Tokyo, Japan.
Oishi
M
M
D000818:Animals; D016590:Aphidicolin; D002499:Centrifugation, Density Gradient; D003513:Cycloheximide / Q000494:pharmacology; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004270:DNA, Circular / Q000096:biosynthesis* / Q000378:metabolism; D004224:Diterpenes / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005111:Extrachromosomal Inheritance; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D013045:Species Specificity; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism*
D004270:DNA, Circular; D004224:Diterpenes; D016590:Aphidicolin; D003513:Cycloheximide; D004257:DNA Polymerase II; D004262:DNA Restriction Enzymes