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PMID: 2841465 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Interaction between the octamer-binding protein nuclear factor III and the adenovirus origin of DNA replication.

Journal of virology ·Vol. 62 ·No. 9 ·1988-09-00 ·Pages 3092-102

Pruijn GJ, van Miltenburg RT, Claessens JA, van der Vliet PC

Abstract

Nuclear factor III (NFIII) is a HeLa sequence-specific DNA-binding protein that stimulates initiation of adenovirus DNA replication in vitro and may be involved in regulation of transcription of several cellular and viral genes. We have studied the interaction between NFIII and the binding site in the adenovirus type 2 (Ad2) origin in detail by methidiumpropyl-EDTA.iron(II) and hydroxyl radical footprinting and by alkylation interference experiments. Our results indicate that (i) the core of the recognition sequence is 5'-TATGATAAT-3'; (ii) both major and minor groove base contacts are detected, and all base pairs in the core are involved in binding; (iii) many backbone contacts are observed divided into a large domain coinciding with the core and a small domain; (iv) contact points are not confined to one side of the DNA helix in contrast to the nuclear factor I (NFI)-binding site; (v) the binding site overlaps the NFI-binding site for at least one nucleotide. A number of Ad2 mutants as well as related binding sites in the origins of other adenovirus serotypes were systematically compared for binding with NFIII. The results are in good agreement with the contact point studies and show that at least one AT base pair is commonly required by NFI and NFIII for optimal binding. The strongest binding site, which contains the octamer/decanucleotide motif (ATGCAAAT[NA]), was found in the Ad4 origin, which lacks an NFI-binding site. Stimulation of in vitro DNA replication of Ad2, Ad4, and Ad12 by NFIII showed that the maximal level of stimulation is dependent on the affinity of NFIII for the origin.

MeSH Terms
Adenoviruses, Human/genetics,physiology Base Sequence Computer Graphics DNA Replication DNA Restriction Enzymes DNA, Viral/metabolism DNA-Binding Proteins/metabolism Densitometry Electrophoresis, Polyacrylamide Gel Genetic Complementation Test HeLa Cells Host Cell Factor C1 Humans Molecular Sequence Data Mutation Nuclear Proteins/metabolism Octamer Transcription Factor-1 Transcription Factors Virus Replication
Chemicals
DNA, Viral DNA-Binding Proteins HCFC1 protein, human Host Cell Factor C1 Nuclear Proteins Octamer Transcription Factor-1 POU2F1 protein, human Transcription Factors DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Pruijn G J
Laboratory for Physiological Chemistry, State University of Utrecht, The Netherlands.
van Miltenburg R T
Claessens J A
van der Vliet P C
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.62.9.3092-3102.1988
Published
1988-09-00
Pages
3092-102
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC253425
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.62.9.3092-3102.1988 PMC Full Text View journal details
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