Different regions of primase subunit p48 control mouse polyomavirus and simian virus 40 DNA replication in vitro.
DNA polymerase alpha-primase (pol-prim), a complex consisting of four subunits, is the major species-specific factor for mouse polyomavirus (PyV) and simian virus 40 (SV40) DNA replication. Although p48 is the most conserved subunit of pol-prim, it is required for in vitro PyV DNA replication but can inhibit cell-free SV40 DNA replication. Production of chimeric human-mouse p48 revealed that different regions of p48 are involved in supporting PyV DNA replication and inhibiting SV40 DNA replication. The N and C-terminal parts of p48 do not have species-specific functions in cell-free PyV DNA replication, but the central part (amino acids [aa] 129 to 320) controls PyV DNA replication in vitro. However, PyV T antigen physically binds to mouse, human, and chimeric pol-prim complexes independently, whether they support PyV DNA replication or not. In contrast to the PyV system, the inhibitory effects of mouse p48 on SV40 DNA replication are mediated by N- and C-terminal regions of p48. Thus, a chimeric p48 containing human aa 1 to 128, mouse aa 129 to 320, and human aa 321 to 418 is active in both PyV and SV40 DNA replication in vitro.
Kautz
A R
AR
Abteilung Biochemie, Institut für Molekulare Biotechnologie e.V., D-07745 Jena, Germany.
Schneider
A
A
Weisshart
K
K
Geiger
C
C
Nasheuer
H P
HP
D000818:Animals; D002478:Cells, Cultured; D004256:DNA Polymerase I / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019915:DNA Primase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D015967:Gene Expression Regulation, Viral; D006801:Humans; D051379:Mice; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D014779:Virus Replication
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D019915:DNA Primase; C116932:DNA polymerase alpha-primase; D004256:DNA Polymerase I