Interdigitated residues within a small region of VP16 interact with Oct-1, HCF, and DNA

Lai, J. S., Herr, W. (July 1997) Interdigitated residues within a small region of VP16 interact with Oct-1, HCF, and DNA. Molecular and Cellular Biology, 17 (7). pp. 3937-46. ISSN 0270-7306 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/9199328

Abstract

Upon infection, the herpes simplex virus (HSV) activator of immediate-early (IE) gene transcription VP16 forms a multiprotein-DNA complex with two cellular proteins, Oct-1 and HCF. First, VP16 associates with HCF independently of DNA, and this association stimulates subsequent association with Oct-1 on the DNA target of VP16 activation, the TAATGARAT motif found in HSV IE promoters. We have analyzed the involvement of VP16 residues lying near the carboxy-terminal transcriptional activation domain of VP16 in associating with HCF, Oct-1, and DNA. To assay VP16 association with HCF, we developed an electrophoretic mobility retardation assay in which HCF is used to retard the mobility of a hybrid VP16-GAL4 DNA-binding domain fusion protein bound to a GAL4 DNA-binding site. Analysis of an extensive set of individual and combined alanine substitutions over a 61-amino-acid region of VP16 shows that, even within a region as small as 13 amino acids, there are separate residues involved in association with either HCF, DNA, or Oct-1 bound to DNA; indeed, of two immediately adjacent amino acids in VP16, one is important for DNA binding and the other is important for HCF binding. These results suggest that a small region in VP16 is important for linking in close juxtaposition the four components of the VP16-induced complex and support the hypothesis that the structure of the Oct-1-VP16 interaction in this complex is similar to that formed by the yeast transcriptional regulatory proteins MATa1 and MAT alpha2. We propose that HCF stabilizes this Oct-1-VP16 interaction.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Binding Sites DNA/ chemistry/metabolism DNA-Binding Proteins/ chemistry/metabolism Hela Cells Herpes Simplex Virus Protein Vmw65/ chemistry/metabolism Host Cell Factor C1 Humans Macromolecular Substances Molecular Sequence Data Multiprotein Complexes Mutagenesis, Site-Directed Octamer Transcription Factor-1 Proteins/ chemistry/metabolism Recombinant Proteins Research Support, U.S. Gov't, P.H.S. Structure-Activity Relationship Transcription Factors/ chemistry/metabolism Transcription, Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein
organism description > virus > herpes simplex virus
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations > mutagenesis
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor
CSHL Authors:
Communities: CSHL labs > Herr lab
Depositing User: Kathleen Darby
Date: July 1997
Date Deposited: 07 May 2014 19:47
Last Modified: 07 May 2014 19:47
PMCID: PMC232246
Related URLs:
URI: http://repository.cshl.edu/id/eprint/29994

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