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Title: N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein

Abstract

The major herpes simplex virus DNA-binding protein, designated ICP8, binds tightly to single-stranded DNA and is required for replication of viral DNA. The sensitivity of the DNA-binding activity of ICP8 to the action of the sulfhydryl reagent N-ethylmaleimide has been examined by using nitrocellulose filter-binding and agarose gel electrophoresis assays. Incubation of ICP8 with N-ethylmaleimide results in a rapid loss of DNA-binding activity. Preincubation of ICP8 with single-stranded DNA markedly inhibits this loss of binding activity. These results imply that a free sulfhydryl group is involved in the interaction of ICP8 with single-stranded DNA and that this sulfhydryl group becomes less accessible to the environment upon binding. Agarose gel electrophoretic analysis of the binding interaction in the presence and absence of N-ethylmaleimide indicates that the cooperative binding exhibited by ICP8 is lost upon treatment with this reagent but that some residual noncooperative binding may remain. This last result was confirmed by equilibrium dialysis experiments with the {sup 32}P-labeled oligonucleotide dT{sub 10} and native and N-ethylmaleimide-treated ICP8.

Authors:
 [1]
  1. Uniformed Services Univ. of the Health Sciences, Bethesda, MD (USA)
Publication Date:
OSTI Identifier:
6531577
Resource Type:
Journal Article
Journal Name:
Journal of Virology; (USA)
Additional Journal Information:
Journal Volume: 62:3; Journal ID: ISSN 0022-538X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; DNA; CROSS-LINKING; NEM; BIOLOGICAL EFFECTS; PROTEINS; VIRUSES; DNA REPLICATION; ELECTROPHORESIS; HERPES SIMPLEX; OLIGONUCLEOTIDES; TRITIUM COMPOUNDS; ANTIMITOTIC DRUGS; CHEMICAL REACTIONS; DISEASES; DRUGS; HYDROGEN COMPOUNDS; IMIDES; INFECTIOUS DISEASES; MICROORGANISMS; NUCLEIC ACID REPLICATION; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PARASITES; POLYMERIZATION; RADIOSENSITIZERS; SKIN DISEASES; VIRAL DISEASES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Ruyechan, W T. N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein. United States: N. p., 1988. Web.
Ruyechan, W T. N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein. United States.
Ruyechan, W T. 1988. "N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein". United States.
@article{osti_6531577,
title = {N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein},
author = {Ruyechan, W T},
abstractNote = {The major herpes simplex virus DNA-binding protein, designated ICP8, binds tightly to single-stranded DNA and is required for replication of viral DNA. The sensitivity of the DNA-binding activity of ICP8 to the action of the sulfhydryl reagent N-ethylmaleimide has been examined by using nitrocellulose filter-binding and agarose gel electrophoresis assays. Incubation of ICP8 with N-ethylmaleimide results in a rapid loss of DNA-binding activity. Preincubation of ICP8 with single-stranded DNA markedly inhibits this loss of binding activity. These results imply that a free sulfhydryl group is involved in the interaction of ICP8 with single-stranded DNA and that this sulfhydryl group becomes less accessible to the environment upon binding. Agarose gel electrophoretic analysis of the binding interaction in the presence and absence of N-ethylmaleimide indicates that the cooperative binding exhibited by ICP8 is lost upon treatment with this reagent but that some residual noncooperative binding may remain. This last result was confirmed by equilibrium dialysis experiments with the {sup 32}P-labeled oligonucleotide dT{sub 10} and native and N-ethylmaleimide-treated ICP8.},
doi = {},
url = {https://www.osti.gov/biblio/6531577}, journal = {Journal of Virology; (USA)},
issn = {0022-538X},
number = ,
volume = 62:3,
place = {United States},
year = {Tue Mar 01 00:00:00 EST 1988},
month = {Tue Mar 01 00:00:00 EST 1988}
}