In situ studies on the effect of acid extraction on the DNA template activity of mature avian erythrocytes.
Exogenous E. coli RNA polymerase was used to determine the in situ DNA template activity of ethanol/acetone fixed avian erythrocytes. No RNA polymerase-catalyzed incorporation of 3H-UTP was detected in mature avian erythrocytes while simultaneously fixed avian lymphocytes did exhibit incorporation of 3H-UTP. Nuclei of mature erythrocytes which were subjected to treatments known to remove histones showed dramatic increases in RNA polymerase-catalyzed incorporation of 3H-UTP. The chromatin of treated cells was presumed to be more accessible to RNA polymerase as determined by the increase in RNA polymerase-catalyzed incorporation of 3H-UTP. Incubation of acid-treated nuclei in poly-L-lysine prior to incubation with RNA polymerase failed to inhibit the incorporation of 3H-UTP. Possible mechanisms for the inactivation of avian erythrocyte nuclei are discussed.
D000818:Animals; D002645:Chickens; D004247:DNA / Q000097:blood* / Q000302:isolation & purification; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003851:Deoxyribonucleases; D004912:Erythrocytes / Q000032:analysis* / Q000648:ultrastructure; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008239:Lysine; D008858:Microscopy, Phase-Contrast; D010455:Peptides / Q000494:pharmacology; D012260:Ribonucleases; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000187:drug effects
D010455:Peptides; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D003851:Deoxyribonucleases; D012260:Ribonucleases; D008239:Lysine