|
|
|
|||||||
|
|||||||||
J Biol Chem, Vol. 275, Issue 4, 2931-2937, January 28, 2000
From the Department of Biochemistry and Molecular Biology,
University of Miami School of Medicine, Miami, Florida 33101-6129
Herpes simplex virus type-1 origin-binding
protein (UL9 protein) initiates viral replication by unwinding the
origins. It possesses sequence-specific DNA-binding activity,
single-stranded DNA-binding activity, DNA helicase activity, and ATPase
activity that is strongly stimulated by single-stranded DNA. We have
examined the role of cysteines in its action as a DNA helicase. The DNA helicase and DNA-dependent ATPase activities of UL9 protein
were stimulated by reducing agent and specifically inactivated by the sulfhydryl-specific reagent N-ethylmaleimide. To identify
the cysteine responsible for this phenomenon, a conserved cysteine in
the vicinity of the ATP-binding site (cysteine 111) was mutagenized to
alanine. UL9C111A protein exhibits defects in its DNA helicase and
DNA-dependent ATPase activities and was unable to support origin-specific DNA replication in vivo. A kinetic analysis
indicates that these defects are due to the inability of
single-stranded DNA to induce high affinity ATP binding in UL9C111A
protein. The DNA-dependent ATPase activity of UL9C111A
protein is resistant to N-ethylmaleimide, while its DNA
helicase activity remains sensitive. Accordingly, sensitivity of UL9
protein to N-ethylmaleimide is due to at least two
cysteines. Cysteine 111 is involved in coupling single-stranded DNA
binding to ATP-binding and subsequent hydrolysis, while a second
cysteine is involved in coupling ATP hydrolysis to DNA unwinding.
Cysteine 111 Affects Coupling of Single-stranded DNA Binding to
ATP Hydrolysis in the Herpes Simplex Virus Type-1 Origin-binding
Protein*
*
This work was supported by National Institutes of Health
Grant AI38335.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Biochemistry
and Molecular Biology, University of Miami School of Medicine, P.O. Box
016129, Miami, FL 33101-6129. Tel.: 305-243-2934; Fax: 305-243-3955;
E-mail: pboehmer@molbio.med.miami.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  A. V. Nimonkar and P. E. Boehmer Role of Protein-Protein Interactions during Herpes Simplex Virus Type 1 Recombination-dependent Replication J. Biol. Chem., May 21, 2004; 279(21): 21957 - 21965. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Nimonkar and P. E. Boehmer In Vitro Strand Exchange Promoted by the Herpes Simplex Virus Type-1 Single Strand DNA-binding Protein (ICP8) and DNA Helicase-Primase J. Biol. Chem., April 19, 2002; 277(17): 15182 - 15189. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Tanguy Le Gac and P. E. Boehmer Activation of the Herpes Simplex Virus Type-1 Origin-binding Protein (UL9) by Heat Shock Proteins J. Biol. Chem., February 8, 2002; 277(7): 5660 - 5666. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. E. Arana, B. Haq, N. Tanguy Le Gac, and P. E. Boehmer Modulation of the Herpes Simplex Virus Type-1 UL9 DNA Helicase by Its Cognate Single-strand DNA-binding Protein, ICP8 J. Biol. Chem., February 23, 2001; 276(9): 6840 - 6845. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |