Species-specific functional interactions of DNA polymerase alpha-primase with simian virus 40 (SV40) T antigen require SV40 origin DNA.
Physical and functional interactions of simian virus 40 (SV40) and polyomavirus large-T antigens with DNA polymerase alpha-primase were analyzed to elucidate the molecular basis for the species specificity of polymerase alpha-primase in viral DNA replication. SV40 T antigen associated more efficiently with polymerase alpha-primase in crude human extracts than in mouse extracts, while polyomavirus T antigen interacted preferentially with polymerase alpha-primase in mouse extracts. The apparent species specificity of complex formation was not observed when purified polymerase alpha-primases were substituted for the crude extracts. Several functional interactions between T antigen and purified polymerase alpha-primase, including stimulation of primer synthesis and primer elongation on M13 DNA in the presence or absence of the single-stranded DNA binding protein RP-A, also proved to be independent of the species from which polymerase alpha-primase had been purified. However, the human DNA polymerase alpha-primase was specifically required for primosome assembly and primer synthesis on SV40 origin DNA in the presence of T antigen and RP-A.
Schneider
C
C
Institute for Biochemistry, Munich, Germany.
Weisshart
K
K
Guarino
L A
LA
Dornreiter
I
I
Fanning
E
E
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D019915:DNA Primase; D017931:DNA Primers; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D013950:Thymus Gland / Q000201:enzymology; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D017931:DNA Primers; D004279:DNA, Viral; D011994:Recombinant Proteins; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases