Search
Filter Results
- Resource Type
- Article1
- Journal1
- Journal Digital1
- Article Type
- Research Support, U.S. Gov't, Non-P.H.S.1
- Research Support, U.S. Gov't, P.H.S.1
- Result From
- Lane Catalog1
- PubMed1
-
Year
- Journal Title
- Proc Natl Acad Sci U S A1
Search Results
Sort by
- JournalDigital AccessProviderVersionWileyCINAHLv. 1-, 1990- Full text delayed 12 monthsAcad Search Premv. 14-, 2003- Full text delayed 12 months
- ArticleScott JF, Eisenberg S, Bertsch LL, Kornberg A.Proc Natl Acad Sci U S A. 1977 Jan;74(1):193-7.The enzyme system for duplicating the duplex, circular DNA of phage phi X174 (replicative form) in stage II of the replicative life cycle was shown to proceed in two steps: synthesis of the viral (+) strand ]stage II(+)], followed by synthesis of the complementary (-) strand ]stage II(-)] [Eisenberg et al. (1976) Proc. Natl. Acad. Sci. USA 73, 3151-3155]. Novel features of the mechanism of the stage II(+) reaction have now been observed. The product, synthesized in extensive net quantities, is a covalently closed, circular, single-stranded DNA. The supercoiled replicative form I template and three of the four required proteins--the phage-induced cistron A protein (cis A), the host rep protein (rep), and the DNA polymerase III holoenzyme (holoenzyme)--act catalytically; the Escherichia coli DNA unwinding (or binding) protein binds the product stoichiometrically. In a reaction uncoupled from replication, cis A, rep, DNA binding protein, ATP, and Mg2+ separate the supercoiled replicative form I into its component single strands coated with DNA binding protein. In the presence of Mg2+, cis A, nicks the replicative form I; rep, ATP, and Mg2+ achieve strand separation with a concurrent cleavage of ATP and binding of DNA binding protein to the single strands. rep exhibits a single-stranded DNA-dependent ATPase activity. These observations suggest that the rep enzymatically melts the duplex at the replicating fork, using energy provided by ATP; this mechanism may apply to the replication of the E. coli chromosome as well.