| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print October 22, 2001
J. Biol. Chem, 10.1074/jbc.M108443200
Submitted on August 31, 2001
Revised on October 17, 2001
Accepted on October 17, 2001
Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115
Corresponding Author: s-j_lee{at}hms.harvard.edu
At a replication fork DNA primase synthesizes oligoribonucleotides which serve as primers for the lagging strand DNA polymerase. In the bacteriophage T7 replication system, DNA primase is encoded by gene 4 of the phage. The 63-kDa gene 4 protein is composed of two major domains, a helicase domain and a primase domain located in the carboxyl and amino terminal halves of the protein, respectively. T7 DNA primase recognizes the sequence 5’-NNGTC-3’ via a zinc motif and catalyzes the template-directed synthesis of tetraribonucleotides pppACNN. T7 DNA primase, like other primases, shares limited homology with DNA-dependent RNA polymerases. In order to identify the catalytic core of the T7 DNA primase, single point mutations were introduced into a basic region that shares sequence homology with RNA polymerases. The genetically altered gene 4 proteins were examined for their ability to support phage growth, to synthesize functional primers, and to recognize primase recognition sites. Two lysine residues, K122 and K128, are essential for phage growth. The two residues play a key role in the synthesis of phosphodiester bonds but are not involved in other activities mediated by the protein. The altered primases are unable to either synthesize or extend an oligoribonucleotide. However, the altered primases do recognize the primase recognition sequence, anneal an exogenous primer 5’-ACCC-3' at the site, and transfer the primer to T7 DNA polymerase. Other lysines in the vicinity are not essential for the synthesis of primers.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S.-J. Lee, U. Qimron, and C. C. Richardson Communication between subunits critical to DNA binding by hexameric helicase of bacteriophage T7 PNAS, July 1, 2008; 105(26): 8908 - 8913. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. Donmez and S. S. Patel Mechanisms of a ring shaped helicase Nucleic Acids Res., September 10, 2006; 34(15): 4216 - 4224. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-J. Lee, B. Marintcheva, S. M. Hamdan, and C. C. Richardson The C-terminal Residues of Bacteriophage T7 Gene 4 Helicase-Primase Coordinate Helicase and DNA Polymerase Activities J. Biol. Chem., September 1, 2006; 281(35): 25841 - 25849. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Fien and J. Hurwitz Fission Yeast Mcm10p Contains Primase Activity J. Biol. Chem., August 4, 2006; 281(31): 22248 - 22260. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Lee and C. C. Richardson Acidic Residues in the Nucleotide-binding Site of the Bacteriophage T7 DNA Primase J. Biol. Chem., July 22, 2005; 280(29): 26984 - 26991. [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]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 S. Vashee, C. Cvetic, W. Lu, P. Simancek, T. J. Kelly, and J. C. Walter Sequence-independent DNA binding and replication initiation by the human origin recognition complex Genes & Dev., August 1, 2003; 17(15): 1894 - 1908. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-J. Lee and C. C. Richardson Interaction of adjacent primase domains within the hexameric gene 4 helicase-primase of bacteriophage T7 PNAS, October 1, 2002; 99(20): 12703 - 12708. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
