| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print March 28, 2001
Department of Biological Chemistry & Molecular Pharmacology, Harvard Medical School, Boston, MA 02115
Corresponding Author: tome{at}hms.harvard.edu
The lagging strand of the replication fork is initially copied as short Okazaki fragments produced by the coupled activities of two template-dependent enzymes, a primase that synthesizes RNA primers and a DNA polymerase that elongates them. Gene 4 of bacteriophage T7 encodes a bifunctional primase-helicase that assembles into a ring-shaped hexamer with both DNA unwinding and primer synthesis activities. The primase is also required for the utilization of RNA primers by T7 DNA polymerase. It is not known how many subunits of the primase-helicase hexamer participate directly in the priming of DNA synthesis. In order to determine the minimal requirements for RNA primer utilization by T7 DNA polymerase, we created an altered gene 4 protein that does not form functional hexamers and consequently lacks detectable DNA unwinding activity. Remarkably, this monomeric primase readily primes DNA synthesis by T7 DNA polymerase on single-stranded templates. The monomeric gene 4 protein forms a specific and stable complex with T7 DNA polymerase and thereby delivers the RNA primer to the polymerase for the onset of DNA synthesis. These results show that a single subunit of the primase-helicase hexamer contains all of the residues required for primer synthesis and for utilization of primers by T7 DNA polymerase.
J. Biol. Chem, 10.1074/jbc.M101470200
Submitted on February 15, 2001
Revised on March 28, 2001
Accepted on March 28, 2001
A complex of the bacteriophage T7 primase-helicase and DNA polymerase directs primer utilization
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Choe, H. H. Guo, and G. van den Engh A dual-fluorescence reporter system for high-throughput clone characterization and selection by cell sorting Nucleic Acids Res., March 14, 2005; 33(5): e49 - e49. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Zhang, M. M. Spiering, M. A. Trakselis, F. T. Ishmael, J. Xi, S. J. Benkovic, and G. G. Hammes Assembly of the bacteriophage T4 primosome: Single-molecule and ensemble studies PNAS, March 1, 2005; 102(9): 3254 - 3259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kato, T. Ito, G. Wagner, and T. Ellenberger A Molecular Handoff between Bacteriophage T7 DNA Primase and T7 DNA Polymerase Initiates DNA Synthesis J. Biol. Chem., July 16, 2004; 279(29): 30554 - 30562. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. K. Levin, Y.-H. Wang, and S. S. Patel The Functional Interaction of the Hepatitis C Virus Helicase Molecules Is Responsible for Unwinding Processivity J. Biol. Chem., June 18, 2004; 279(25): 26005 - 26012. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Lee and C. C. Richardson The Linker Region between the Helicase and Primase Domains of the Gene 4 Protein of Bacteriophage T7: ROLE IN HELICASE CONFORMATION AND ACTIVITY J. Biol. Chem., May 28, 2004; 279(22): 23384 - 23393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-G. He and C. C. Richardson Effect of Single-stranded DNA-binding Proteins on the Helicase and Primase Activities of the Bacteriophage T7 Gene 4 Protein J. Biol. Chem., May 21, 2004; 279(21): 22190 - 22197. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. M. Miller, C. M. Gouvion, B. L. Davidson, and H. L. Paulson Targeting Alzheimer's disease genes with RNA interference: an efficient strategy for silencing mutant alleles Nucleic Acids Res., January 30, 2004; 32(2): 661 - 668. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Lee and C. C. Richardson Essential Lysine Residues in the RNA Polymerase Domain of the Gene 4 Primase-Helicase of Bacteriophage T7 J. Biol. Chem., December 21, 2001; 276(52): 49419 - 49426. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
