Effects of in vitro dephosphorylation on DNA-binding and DNA helicase activities of simian virus 40 large tumor antigen.
Simian virus 40 large T antigen is a phosphoprotein with two clusters of phosphorylation sites. Each cluster includes four serine residues and one threonine residue. In vitro treatment with intestinal alkaline phosphatase removes the phosphate groups from the serine but not from the threonine residues. Potato acid phosphatase additionally dephosphorylates the phosphothreonine (Thr-124) in the N-terminal cluster but does not attack the phosphothreonine in the C-terminal cluster (Thr-701). Two biochemical functions of untreated and partially dephosphorylated T antigen were assayed, namely, its specific DNA-binding property and its DNA helicase activity. After treatment with alkaline phosphatase, T antigen had a severalfold higher affinity for the specific binding sites in the viral genomic control region, in particular, for binding site II in the origin of replication. However, T antigen, when dephosphorylated by acid phosphatase, had DNA-binding properties similar to those of the untreated control. Neither alkaline nor acid dephosphorylation affected the DNA helicase activity of T antigen.
Klausing
K
K
Fakultät für Biologie, Universität Konstanz, Federal Republic of Germany.
Scheidtmann
K H
KH
Baumann
E A
EA
Knippers
R
R
D000135:Acid Phosphatase / Q000494:pharmacology; D000469:Alkaline Phosphatase / Q000494:pharmacology; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism* / Q000502:physiology; D001665:Binding Sites; D002460:Cell Line; D004265:DNA Helicases / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins; D010449:Peptide Mapping; D010766:Phosphorylation; D013539:Simian virus 40 / Q000201:enzymology / Q000235:genetics / Q000276:immunology*
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D029968:Escherichia coli Proteins; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; C027093:TraI protein, E coli; D004265:DNA Helicases