Studies of Schizosaccharomyces pombe DNA polymerase alpha at different stages of the cell cycle.
The status of Schizosaccharomyces pombe (fission yeast) DNA polymerase alpha was investigated at different stages of the cell cycle. S.pombe DNA polymerase alpha is a phosphoprotein, with serine being the exclusive phosphoamino acid. By in vivo pulse labeling experiments DNA polymerase alpha was found to be phosphorylated to a 3-fold higher level in late S phase cells compared with cells in the G2 and M phases, but the steady-state level of phosphorylation did not vary significantly during the cell cycle. Tryptic phosphopeptide mapping demonstrated that the phosphorylation sites of DNA polymerase alpha from late S phase cells were not the same as that from G2/M phase cells. DNA polymerase alpha partially purified from G1/S cells had a different mobility in native gels from that from G2/M phase cells. The partially purified polymerase alpha from G1/S phase cells had a higher affinity for single-stranded DNA than that from G2/M phase cells. Despite the apparent differences in cell cycle-dependent phosphorylation, mobility in native gels and affinity for DNA, the in vitro enzymatic activity of the partially purified DNA polymerase alpha did not appear to vary during the cell cycle. The possible biological significance of these cell cycle-dependent characteristics of DNA polymerase alpha is discussed.
Park
H
H
Department of Pathology, Stanford University School of Medicine, CA 94305, USA.
Davis
R
R
Wang
T S
TS
D002453:Cell Cycle / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D004257:DNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004586:Electrophoresis; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D010449:Peptide Mapping; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000254:growth & development; D020687:cdc25 Phosphatases
D018797:Cell Cycle Proteins; D004277:DNA, Single-Stranded; D010750:Phosphoproteins; D004257:DNA Polymerase II; D010749:Phosphoprotein Phosphatases; D020687:cdc25 Phosphatases