Nucleotide polymerases in the developing avian erythrocyte.
Avian erythroid cells were separated into five developmental stages by sedimentation on discontinuous isotonic albumin gradients. Solubilized enzyme activities from whole cells were partially purified and characterized by ion exchange and ion filtration chromatography and velocity sedimenttation analysis. Three nucleotide polymerase types were investigated: (a) DNA-dependent RNA polymerases; (b) RNA-dependent terminal ribonucleotidyltransferases, and (c) DNA-dependent DNA polymerases. The two characteristic forms of eucaryotic DNA-dependent RNA polymerases, polymerase I (nucleolar) and polymerase II (nucleoplasmic), were identified. Polymerase III was only marginally detectable even in the earliest developmental populations. At least two species of RNA-dependent terminal ribosyltransferases were present. One apparently was the poly(A) polymerase observed in other systems. The other terminal transferase was present in two chromatographic forms, required an RNA primer, and used UTP and/or CTP as particularly efficient substrates. Three DNA polymerase activities were resolved, two of which were characteristic of the alpha and beta DNA polymerases described in other eucaryotic systems. The third polymerase was not the gamma polymerase but a separate entity. Poly(dC)-dependent RNA polymerase activity, associated with the alpha polymerase, was relatively enriched in the third DNA polymerase species. The activity levels of the nucleotide polymerases were monitored as a function of red cell maturation. Characteristic declining patterns of activity were obtained for each enzyme which correlate well with the synthetic rates of their in vivo products where these are known. These results correlate well with the synthetic rates of their in vivo products where these are known. These results are consistent with the postulate that the general transcriptive and replicative control processes operating during development may involve changes in the level of the requisite polymerases.
D000740:Anemia / Q000201:enzymology; D000818:Animals; D001853:Bone Marrow / Q000201:enzymology; D001854:Bone Marrow Cells; D002645:Chickens; D004256:DNA Polymerase I / Q000097:blood* / Q000302:isolation & purification; D004257:DNA Polymerase II / Q000097:blood* / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase / Q000097:blood*; D012321:DNA-Directed RNA Polymerases / Q000097:blood* / Q000302:isolation & purification; D004900:Erythroblasts / Q000201:enzymology / Q000648:ultrastructure; D004912:Erythrocytes / Q000201:enzymology* / Q000648:ultrastructure; D005260:Female
D012321:DNA-Directed RNA Polymerases; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase