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Science 16 September 1988:
Vol. 241. no. 4872, pp. 1486 - 1489
DOI: 10.1126/science.2843984
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Articles

Science, Vol 241, Issue 4872, 1486-1489
Copyright © 1988 by American Association for the Advancement of Science

articles

An origin unwinding activity regulates initiation of DNA replication during mammalian cell cycle

JM Roberts and G D'Urso

Fred Hutchinson Cancer Research Center, Seattle, WA 98104.

An in vitro assay was developed to study the positive factors that regulate the onset of DNA replication during the mammalian cell cycle. Extracts prepared from cells at defined positions in the cell cycle were used to examine the replication of SV40 DNA in a cell free system. Extracts prepared from S phase cells were ten times more efficient at initiating replication at the SV40 origin than were extracts from G1 cells, whereas elongation rates were similar in G1 and S reactions. At a discrete point in the cell cycle, just before the cell's entry into S, an activity appeared that was required, in conjunction with SV40 T antigen, for site specific initiation at the SV40 origin. This factor had a role in unwinding DNA at the replication origin.


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S phase of the cell cycle.
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