Yeast replication factor-A functions in the unwinding of the SV40 origin of DNA replication

TY - JOUR

T1 - Yeast replication factor-A functions in the unwinding of the SV40 origin of DNA replication

AU - Brill, Steven J.

AU - Stillman, Bruce

PY - 1989

Y1 - 1989

N2 - CELL-free replication systems for simian virus 40 (SV40) DNA are taken to be a model for the replication of eukaryotic chromosomes, because only one viral protein is required to supplement the replication proteins provided by a human cell extract. To prove that these cellular proteins function in chromosomal DNA replication we have begun to identify homologous proteins in an organism that can be genetically manipulated. Here we report the identification of yeast replication factor-A (yRF-A) from Saccharomyces cerevisiae and show that it is functionally and structurally related to a human protein that is required for the initiation and elongation of SV40 DNA replication. Yeast RF-A, a multi-subunit phosphoprotein, is similar to the human protein in its chromatographic behaviour, subunit structure and DNA-binding activity. The yeast protein will fully substitute for the human protein in an early stage of the initiation of SV40 DNA replication. Substitution of yRF-A in the complete SV40 replication system, however, results in reduced DNA replication, presumably due to a require-ment for species-specific interactions between yeast RF-A and the DNA polymerase complex.

AB - CELL-free replication systems for simian virus 40 (SV40) DNA are taken to be a model for the replication of eukaryotic chromosomes, because only one viral protein is required to supplement the replication proteins provided by a human cell extract. To prove that these cellular proteins function in chromosomal DNA replication we have begun to identify homologous proteins in an organism that can be genetically manipulated. Here we report the identification of yeast replication factor-A (yRF-A) from Saccharomyces cerevisiae and show that it is functionally and structurally related to a human protein that is required for the initiation and elongation of SV40 DNA replication. Yeast RF-A, a multi-subunit phosphoprotein, is similar to the human protein in its chromatographic behaviour, subunit structure and DNA-binding activity. The yeast protein will fully substitute for the human protein in an early stage of the initiation of SV40 DNA replication. Substitution of yRF-A in the complete SV40 replication system, however, results in reduced DNA replication, presumably due to a require-ment for species-specific interactions between yeast RF-A and the DNA polymerase complex.

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U2 - https://doi.org/10.1038/342092a0

DO - https://doi.org/10.1038/342092a0

M3 - Article

C2 - 2554144

VL - 342

SP - 92

EP - 95

JO - Nature

JF - Nature

SN - 0028-0836

IS - 6245

ER -