| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Department of Virology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-4000, U.S.A.
Herpes simplex virus type 1 encodes its own DNA polymerase (Pol), the product of the UL30 gene, and a polymerase accessory subunit, the product of the UL42 gene, both of which are required for viral DNA replication. Pol and the UL42 protein associate to form a heterodimeric complex (Pol/UL42) which is more active and has a higher processivity than the Pol catalytic subunit alone. The Pol/UL42 complex has been reconstituted by mixing together highly purified Pol and UL42 subunits obtained from recombinant baculovirus-infected cells. We have used polymerase activity on poly(dA):oligo(dT20), a template that the Pol subunit utilizes with low efficiency, to measure the formation of the Pol/UL42 complex. Our data indicate that the association constant for the Pol/UL42 complex is 1 x 108 M-1. Proteolytic digestions of UL42 were performed to determine whether structural domains of UL42 could be disclosed by differential amino acid accessibilities. The ability of these protease-resistant domains to form a functional complex with Pol was determined by measuring their ability to stimulate Pol activity on poly(dA):oligo(dT20). We have found that trypsin digestion of UL42 in the presence of DNA generates protease-resistant fragments of 28K and 8K which co-elute from a MonoQ column and are able to stimulate Pol activity on poly(dA):oligo(dT20). Complex formation of the 28K and 8K tryptic fragments with Pol was also shown by their co-immunoprecipitation with antibody to Pol. It was determined that the 28K fragment of UL42 comprised amino acids 1 to 245 or 1 to 254 of UL42, whereas the 8K fragment started at amino acid 255. Thus, controlled proteolysis of UL42 revealed two closely contiguous structural domains that retained the ability to complex with Pol and stimulate Pol activity.
Present address: Department of Biochemistry, Bio-mega/Boehringer Ingelheim Research Inc., Laval, Quebec, Canada.
Received 1 March 1993;
accepted 21 May 1993.
This article has been cited by other articles:
|
|
 |
|
  A. Loregian, A. Case, E. Cancellotti, C. Valente, H. S. Marsden, and G. Palu Cloning, expression, and functional characterization of the equine herpesvirus 1 DNA polymerase and its accessory subunit. J. Virol., July 1, 2006; 80(13): 6247 - 6258. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. W. Randell, G. Komazin, C. Jiang, C. B. C. Hwang, and D. M. Coen Effects of Substitutions of Arginine Residues on the Basic Surface of Herpes Simplex Virus UL42 Support a Role for DNA Binding in Processive DNA Synthesis J. Virol., September 15, 2005; 79(18): 12025 - 12034. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Loregian, B. A. Appleton, J. M. Hogle, and D. M. Coen Residues of Human Cytomegalovirus DNA Polymerase Catalytic Subunit UL54 That Are Necessary and Sufficient for Interaction with the Accessory Protein UL44 J. Virol., January 1, 2004; 78(1): 158 - 167. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. G. Bridges, C. S. Chow, and D. M. Coen Identification of Crucial Hydrogen-Bonding Residues for the Interaction of Herpes Simplex Virus DNA Polymerase Subunits via Peptide Display, Mutational, and Calorimetric Approaches J. Virol., June 1, 2001; 75(11): 4990 - 4998. [Abstract] [Full Text]
|
|
|
|
|
|
S. R. Chan and B. Chandran Characterization of Human Herpesvirus 8 ORF59 Protein (PF-8) and Mapping of the Processivity and Viral DNA Polymerase-Interacting Domains J. Virol., December 1, 2000; 74(23): 10920 - 10929. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. G. Bridges, Q. Hua, M. R. Brigham-Burke, J. D. Martin, P. Hensley, C. E. Dahl, P. Digard, M. A. Weiss, and D. M. Coen Secondary Structure and Structure-Activity Relationships of Peptides Corresponding to the Subunit Interface of Herpes Simplex Virus DNA Polymerase J. Biol. Chem., January 7, 2000; 275(1): 472 - 478. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. R. Lehman and P. E. Boehmer Replication of Herpes Simplex Virus DNA J. Biol. Chem., October 1, 1999; 274(40): 28059 - 28062. [Full Text] [PDF]
|
|
|
|
|
|
D. J. Tenney, A. K. Sheaffer, W. W. Hurlburt, M. Bifano, and R. K. Hamatake Sequence-dependent Primer Synthesis by the Herpes Simplex Virus Helicase-Primase Complex J. Biol. Chem., April 21, 1995; 270(16): 9129 - 9136. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
