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PMID: 2247056 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

RIP60, a mammalian origin-binding protein, enhances DNA bending near the dihydrofolate reductase origin of replication.

Molecular and cellular biology ·Vol. 10 ·No. 12 ·1990-12-00 ·Pages 6236-43

Caddle MS, Dailey L, Heintz NH

Abstract

Replication of the Chinese hamster dihydrofolate (dhfr) gene initiates near a 281-bp HaeIII fragment of stably bent DNA that binds RIP60, a 60-kDa origin-specific DNA-binding protein that has been purified from HeLa cell nuclear extract (L. Dailey, M. S. Caddle, N. Heintz, and N. H. Heintz, Mol. Cell. Biol. 10:6225-6235, 1990). Circular permutation assays showed that stable DNA bending in the dhfr origin region fragment was due to the presence of five oligo (dA)3-4 tracts, designated bend elements B1 to B5, that are spaced 10 bp apart. DNA bending directed by elements B1 to B5, as assessed by anomolous migration of DNA fragments on polyacrylamide gels, was accentuated at 4 degrees C. Bend element B5, which is in inverse orientation relative to elements B1 to B4, overlaps an ATT-rich motif that comprises the RIP60 protein-binding site. Gel mobility shift assays with circularly permuted bent DNA fragments and purified RIP60 showed that RIP60 markedly enhanced DNA bending of the dhfr origin region sequences. These results suggest that, as in many plasmids, bacteriophages, and eucaryotic viruses, mammalian DNA-binding proteins may enhance DNA bending near origins of replication during initiation of DNA synthesis.

Related Genes
dhfr
MeSH Terms
Animals Base Composition Base Sequence Cell Line DNA/genetics,isolation & purification,metabolism DNA Replication DNA-Binding Proteins/metabolism Electrophoresis, Gel, Two-Dimensional Molecular Sequence Data Restriction Mapping Software Tetrahydrofolate Dehydrogenase/genetics Thermodynamics
Chemicals
DNA-Binding Proteins DNA Tetrahydrofolate Dehydrogenase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Caddle M S
Department of Biochemistry, University of Vermont College of Medicine, Burlington 05405.
Dailey L
Heintz N H
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.12.6236-6243.1990
Published
1990-12-00
Pages
6236-43
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362898
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.10.12.6236-6243.1990 PMC Full Text View journal details
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