N-ethylmaleimide inhibition of the DNA-binding activity of the herpes simplex virus type 1 major DNA-binding protein
- Uniformed Services Univ. of the Health Sciences, Bethesda, MD (USA)
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.
- OSTI ID:
- 6531577
- Journal Information:
- Journal of Virology; (USA), Vol. 62:3; ISSN 0022-538X
- Country of Publication:
- United States
- Language:
- English
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DNA
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BIOLOGICAL EFFECTS
PROTEINS
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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
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550201* - Biochemistry- Tracer Techniques