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Identification of cellular components required for SV40 DNA replication in vitro
M Fairman1, G Prelich, T Tsurimoto
1Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724.
Biochimica Et Biophysica Acta
|December 20, 1988
Summary
Researchers identified eight cellular proteins essential for simian virus 40 (SV40) replication, including proliferating-cell nuclear antigen (PCNA), and proposed a multi-stage DNA synthesis model.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- Simian virus 40 (SV40) replication requires specific cellular proteins.
- Understanding these host factors is crucial for elucidating viral DNA replication mechanisms.
Purpose of the Study:
- To identify and characterize cellular proteins involved in SV40 DNA replication.
- To propose a model for SV40 DNA synthesis.
Main Methods:
- Fractionation of human 293 cell extracts.
- Reconstitution assays with viral T antigen (TAg) and SV40 origin DNA.
- Identification of essential protein factors.
Main Results:
- At least eight cellular components are required for SV40 replication.
- Proliferating-cell nuclear antigen (PCNA) is one essential factor.
- Identified replication factors RF-A and RF-C, involved in pre-synthesis complex formation and strand synthesis, respectively.
- Topoisomerases I and II are also necessary for complete replication.
Conclusions:
- SV40 DNA replication involves a complex interplay of multiple cellular proteins.
- A multi-stage model for DNA synthesis, including pre-replication steps, is proposed.