| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 266, Issue 30, 20034-20044, Oct, 1991
MM Munn and BM Alberts
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0448.
We have used DNA footprinting techniques to analyze the interactions of five DNA replication proteins at a primer-template junction: the bacteriophage T4 DNA polymerase (the gene 43 protein), its three accessory proteins (the gene 44/62 and 45 proteins), and the gene 32 protein, which is the T4 helix-destabilizing (or single-stranded DNA- binding) protein. The 177-nucleotide-long DNA substrate consisted of a perfect 52-base pair hairpin helix with a protruding single-stranded 5' tail. As expected, the DNA polymerase binds near the 3' end of this molecule (at the primer-template junction) and protects the adjacent double-stranded region from cleavage. When the gene 32 protein binds to the single-stranded tail, it reduces the concentration of the DNA polymerase required to observe the polymerase footprint by 10-30-fold. Periodic ATP hydrolysis by the 44/62 protein is required to maintain the activity of the DNA polymerase holoenzyme (a complex of the 43, 44/62, and 45 proteins). Footprinting experiments demonstrate the formation of a weak complex between the DNA polymerase and the gene 45 protein, but there is no effect of the 44/62 protein or ATP on this enlarged footprint. We propose a model for holoenzyme function in which the complex of the three accessory proteins uses ATP hydrolysis to keep a moving polymerase tightly bound to the growing 3' end, providing a "clock" to measure polymerase stalling.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Yang, M. A. Trakselis, R. M. Roccasecca, and S. J. Benkovic The Application of a Minicircle Substrate in the Study of the Coordinated T4 DNA Replication J. Biol. Chem., December 12, 2003; 278(50): 49828 - 49838. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Delagoutte and P. H. von Hippel Function and Assembly of the Bacteriophage T4 DNA Replication Complex: INTERACTIONS OF THE T4 POLYMERASE WITH VARIOUS MODEL DNA CONSTRUCTS J. Biol. Chem., July 3, 2003; 278(28): 25435 - 25447. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Wong, G. A. Kassavetis, J.-P. Leonetti, and E. P. Geiduschek Mutational and Functional Analysis of a Segment of the Sigma Family Bacteriophage T4 Late Promoter Recognition Protein gp55 J. Biol. Chem., February 21, 2003; 278(9): 7073 - 7080. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Sun and Y. Shamoo Biochemical Characterization of Interactions between DNA Polymerase and Single-stranded DNA-binding Protein in Bacteriophage RB69 J. Biol. Chem., February 7, 2003; 278(6): 3876 - 3881. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Yao, F. P. Leu, J. Anjelkovic, J. Turner, and M. O'Donnell DNA Structure Requirements for the Escherichia coli gamma Complex Clamp Loader and DNA Polymerase III Holoenzyme J. Biol. Chem., April 6, 2000; 275(15): 11440 - 11450. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. M. Hingorani and M. O'Donnell ATP Binding to the Escherichia coli Clamp Loader Powers Opening of the Ring-shaped Clamp of DNA Polymerase III Holoenzyme J. Biol. Chem., September 18, 1998; 273(38): 24550 - 24563. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tougu and K. J. Marians The Interaction between Helicase and Primase Sets the Replication Fork Clock J. Biol. Chem., August 30, 1996; 271(35): 21398 - 21405. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Herendeen, G. Kassavetis, and E. Geiduschek A transcriptional enhancer whose function imposes a requirement that proteins track along DNA Science, May 29, 1992; 256(5061): 1298 - 1303. [Abstract] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
