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Different regions of primase subunit p48 control mouse polyomavirus and simian virus 40 DNA replication in vitro

A R Kautz1, A Schneider, K Weisshart

  • 1Abteilung Biochemie, Institut für Molekulare Biotechnologie e.V., D-07745 Jena, Germany.

Journal of Virology
|February 13, 2001
PubMed

Insights

The p48 subunit of DNA polymerase alpha-primase plays a key role in polyomavirus DNA replication. Specific regions of p48 differentially regulate replication for mouse polyomavirus and simian virus 40.

Area of Science:

  • Molecular Biology
  • Virology
  • Biochemistry

Background:

  • DNA polymerase alpha-primase (pol-prim) is crucial for viral DNA replication.
  • The p48 subunit is the most conserved component of pol-prim.
  • Species-specific interactions govern polyomavirus DNA replication.

Purpose of the Study:

  • To investigate the species-specific functions of the p48 subunit in polyomavirus DNA replication.
  • To identify regions of p48 responsible for supporting mouse polyomavirus (PyV) and inhibiting simian virus 40 (SV40) DNA replication.

Main Methods:

  • Production and analysis of chimeric human-mouse p48 subunits.
  • In vitro DNA replication assays for PyV and SV40.
  • Analysis of interactions between p48 and viral T antigens.

Main Results:

  • The central region (amino acids 129-320) of p48 controls PyV DNA replication.
  • N- and C-terminal regions of p48 mediate inhibition of SV40 DNA replication.
  • A chimeric p48 subunit (human aa 1-128, mouse aa 129-320, human aa 321-418) demonstrated activity in both PyV and SV40 replication.

Conclusions:

  • The p48 subunit exhibits distinct functional domains for PyV and SV40 DNA replication.
  • Chimeric p48 constructs can restore or enhance replication activity across different polyomaviruses.
  • Understanding these species-specific interactions provides insights into viral replication mechanisms.

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