Physical interaction between the herpes simplex virus 1 origin-binding protein and single-stranded DNA-binding protein ICP8

Paul E Boehmer, I. R. Lehman

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

We had previously demonstrated that the herpes simplex virus 1 (HSV-1) single-stranded DNA-binding protein (ICP8) can specifically stimulate the helicase activity of the HSV-1 origin-binding protein (UL9). We show here that this functional stimulation is a manifestation of a tight interaction between UL9 protein and ICP8. By using protein-affinity chromatography, we have demonstrated the specific binding of purified UL9 protein to immobilized ICP8 and vice versa. Furthermore, ICP8 is specifically retained by a column on which the C-terminal 37-kDa DNA-binding domain of the UL9 protein was immobilized. The interaction between ICP8 and the DNA-binding domain of the UL9 protein was confirmed by cochromatography of the two proteins. These results suggest that the UL9 protein and ICP8 form a tight complex that functions in origin recognition and unwinding.

Original languageEnglish (US)
Pages (from-to)8444-8448
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number18
StatePublished - Sep 15 1993
Externally publishedYes

Fingerprint

DNA-Binding Proteins
Human Herpesvirus 1
Carrier Proteins
Immobilized Proteins
Proteins
DNA
Affinity Chromatography
Protein Domains

Keywords

  • DNA replication
  • Origin unwinding
  • Protein-affinity chromatography
  • Protein-protein interactions

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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abstract = "We had previously demonstrated that the herpes simplex virus 1 (HSV-1) single-stranded DNA-binding protein (ICP8) can specifically stimulate the helicase activity of the HSV-1 origin-binding protein (UL9). We show here that this functional stimulation is a manifestation of a tight interaction between UL9 protein and ICP8. By using protein-affinity chromatography, we have demonstrated the specific binding of purified UL9 protein to immobilized ICP8 and vice versa. Furthermore, ICP8 is specifically retained by a column on which the C-terminal 37-kDa DNA-binding domain of the UL9 protein was immobilized. The interaction between ICP8 and the DNA-binding domain of the UL9 protein was confirmed by cochromatography of the two proteins. These results suggest that the UL9 protein and ICP8 form a tight complex that functions in origin recognition and unwinding.",
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AU - Lehman, I. R.

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N2 - We had previously demonstrated that the herpes simplex virus 1 (HSV-1) single-stranded DNA-binding protein (ICP8) can specifically stimulate the helicase activity of the HSV-1 origin-binding protein (UL9). We show here that this functional stimulation is a manifestation of a tight interaction between UL9 protein and ICP8. By using protein-affinity chromatography, we have demonstrated the specific binding of purified UL9 protein to immobilized ICP8 and vice versa. Furthermore, ICP8 is specifically retained by a column on which the C-terminal 37-kDa DNA-binding domain of the UL9 protein was immobilized. The interaction between ICP8 and the DNA-binding domain of the UL9 protein was confirmed by cochromatography of the two proteins. These results suggest that the UL9 protein and ICP8 form a tight complex that functions in origin recognition and unwinding.

AB - We had previously demonstrated that the herpes simplex virus 1 (HSV-1) single-stranded DNA-binding protein (ICP8) can specifically stimulate the helicase activity of the HSV-1 origin-binding protein (UL9). We show here that this functional stimulation is a manifestation of a tight interaction between UL9 protein and ICP8. By using protein-affinity chromatography, we have demonstrated the specific binding of purified UL9 protein to immobilized ICP8 and vice versa. Furthermore, ICP8 is specifically retained by a column on which the C-terminal 37-kDa DNA-binding domain of the UL9 protein was immobilized. The interaction between ICP8 and the DNA-binding domain of the UL9 protein was confirmed by cochromatography of the two proteins. These results suggest that the UL9 protein and ICP8 form a tight complex that functions in origin recognition and unwinding.

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