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Mouse DNA primase plays the principal role in determination of permissiveness for polyomavirus DNA replication
1Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Abstract:
We have investigated the species-specific replication of polyomavirus DNA in the cell-free system that was established previously (Y. Murakami, T. Eki, M. Yamada, C. Prives, and J. Hurwitz, Proc. Natl. Acad. Sci. USA 83:6347-6351, 1986). Extracts from various species of cells supported polyomavirus DNA replication in a species-specific manner that was consistent with the host range specificity of polyomavirus; extracts prepared from mouse and hamster cells were active, whereas extracts prepared from human, monkey, and insect cells were inactive. The addition of DNA polymerase alpha-primase purified from mouse cells induced the replication of polyomavirus DNA in a cell-free system containing polyomavirus large tumor antigen and nonpermissive cell extracts, such as human and insect cell extracts. Isolated mouse DNA primase alone also induced polyomavirus DNA replication in human cell extracts but not in insect cell extracts, indicating that mouse DNA primase plays the principal role in determining permissiveness for polyomavirus DNA replication.
Insights
Polyomavirus DNA replication is species-specific, with mouse DNA primase being key. This enzyme enables replication in non-permissive human cell extracts, highlighting its crucial role in host range determination.
Area of Science:
- Molecular Biology
- Virology
- Genetics
Background:
- Polyomavirus DNA replication exhibits host range specificity.
- A previously established cell-free system was utilized for investigation.
Purpose of the Study:
- To investigate the species-specific replication of polyomavirus DNA.
- To identify the key cellular factors determining polyomavirus host range.
Main Methods:
- Utilized a cell-free system for polyomavirus DNA replication studies.
- Tested cell extracts from various species (mouse, hamster, human, monkey, insect).
- Assessed the effect of purified mouse DNA polymerase alpha-primase and DNA primase on replication in non-permissive extracts.
Main Results:
- Polyomavirus DNA replication was supported by extracts from permissive species (mouse, hamster) but not non-permissive ones (human, monkey, insect).
- Addition of mouse DNA polymerase alpha-primase restored replication in non-permissive human and insect cell extracts.
- Mouse DNA primase alone induced replication in human cell extracts, but not insect cell extracts.
Conclusions:
- Polyomavirus DNA replication is strictly species-specific.
- Mouse DNA primase is a principal determinant of permissiveness for polyomavirus DNA replication.
- This finding elucidates a key mechanism underlying polyomavirus host range specificity.