Initiation of Bidirectional Replication at the Chromosomal Origin Is Directed by the Interaction between Helicase and Primase*

Several protein-protein interactions have been shown to be critical for proper replication fork function inEscherichia coli. These include interactions between the polymerase and the helicase, the helicase and the primase, and the primase and the polymerase. We have studied the influence of these interactions on proper initiation at oriC by using mutant primases defective in their interaction with the helicase and DNA polymerase III holoenzyme lacking the τ subunit so that it will not interact with the helicase. We show here that accurate initiation of bidirectional DNA replication from oriC is dependent on proper placement of the primers for leading strand synthesis and is thus governed primarily by the interaction between the helicase and primase.

[1]  K. Marians,et al.  The Interaction between Helicase and Primase Sets the Replication Fork Clock* , 1996, The Journal of Biological Chemistry.

[2]  Ronald W. Davis,et al.  SPA1: A gene important for chromosome segregation and other mitotic functions in S. cerevisiae , 1988, Cell.

[3]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[4]  J. Reems,et al.  Coordinated leading- and lagging-strand synthesis at the Escherichia coli DNA replication fork. V. Primase action regulates the cycle of Okazaki fragment synthesis. , 1992, The Journal of biological chemistry.

[5]  S. Kim,et al.  Protects in the Leading-strand Polymerase Complex at the Replication Fork (*) , 1996, The Journal of Biological Chemistry.

[6]  E. Zechner,et al.  Coordinated leading- and lagging-strand synthesis at the Escherichia coli DNA replication fork. II. Frequency of primer synthesis and efficiency of primer utilization control Okazaki fragment size. , 1992, The Journal of biological chemistry.

[7]  T. Baker,et al.  Complete enzymatic replication of plasmids containing the origin of the Escherichia coli chromosome. , 1986, The Journal of biological chemistry.

[8]  W. Soeller,et al.  Maximal limits of the Escherichia coli replication factor Y effector site sequences in pBR322 DNA. , 1982, The Journal of biological chemistry.

[9]  E. Zechner,et al.  Coordinated leading- and lagging-strand synthesis at the Escherichia coli DNA replication fork. III. A polymerase-primase interaction governs primer size. , 1992, The Journal of biological chemistry.

[10]  K. Marians.  Prokaryotic DNA replication. , 1992, Annual review of biochemistry.

[11]  Douglas W. Smith,et al.  DNA replication, the bacterial cell cycle, and cell growth , 1992, Cell.

[12]  E. Zechner,et al.  Coordinated leading- and lagging-strand synthesis at the Escherichia coli DNA replication fork. I. Multiple effectors act to modulate Okazaki fragment size. , 1992, The Journal of biological chemistry.

[13]  H. Hiasa,et al.  Decatenating activity of Escherichia coli DNA gyrase and topoisomerases I and III during oriC and pBR322 DNA replication in vitro. , 1994, The Journal of biological chemistry.

[14]  K. Marians,et al.  Identification of a domain of Escherichia coli primase required for functional interaction with the DnaB helicase at the replication fork. , 1994, The Journal of biological chemistry.

[15]  H. Hiasa,et al.  Primase couples leading- and lagging-strand DNA synthesis from oriC. , 1994, The Journal of biological chemistry.

[16]  H. Hiasa,et al.  DNA Strand Cleavage Is Required for Replication Fork Arrest by a Frozen Topoisomerase-Quinolone-DNA Ternary Complex* , 1996, The Journal of Biological Chemistry.

[17]  C. McHenry,et al.  Coupling of a Replicative Polymerase and Helicase: A τ–DnaB Interaction Mediates Rapid Replication Fork Movement , 1996, Cell.

[18]  K. Marians,et al.  The Extreme C Terminus of Primase Is Required for Interaction with DnaB at the Replication Fork* , 1996, The Journal of Biological Chemistry.

[19]  A. Kornberg,et al.  Replication initiated at the origin (oriC) of the E. coli chromosome reconstituted with purified enzymes , 1984, Cell.

[20]  J. Marszalek,et al.  DnaA protein directs the binding of DnaB protein in initiation of DNA replication in Escherichia coli. , 1994, The Journal of biological chemistry.

[21]  D. Prescott,et al.  Bidirectional replication of the chromosome in Escherichia coli. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[22]  D. Bastia,et al.  Mechanism of recruitment of DnaB helicase to the replication origin of the plasmid pSC101. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[23]  A. Kornberg,et al.  dnaG gene product, a rifampicin-resistant RNA polymerase, initiates the conversion of a single-stranded coliphage DNA to its duplex replicative form. , 1975, The Journal of biological chemistry.

[24]  J. Schweizer,et al.  Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication , 1992, Journal of virology.

[25]  M. O’Donnell,et al.  Replisome Assembly Reveals the Basis for Asymmetric Function in Leading and Lagging Strand Replication , 1996, Cell.

[26]  A. Kornberg,et al.  Replication of phage G4. A novel and simple system for the initiation of deoxyribonucleic acid synthesis. , 1975, The Journal of biological chemistry.

[27]  A. Kornberg,et al.  Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. Bird,et al.  Origin and sequence of chromosome replication in Escherichia coli. , 1972, Journal of molecular biology.

[29]  J. Hurwitz,et al.  The replication of DNA containing the simian virus 40 origin by the monopolymerase and dipolymerase systems. , 1992, Journal of Biological Chemistry.