The origin recognition complex interacts with a bipartite DNA binding site within yeast replicators

Rao, H., Stillman, B. (March 1995) The origin recognition complex interacts with a bipartite DNA binding site within yeast replicators. Proceedings of the National Academy of Sciences of the United States of America, 92 (6). pp. 2224-8. ISSN 0027-8424

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Replicators are genetically defined elements within chromosomes that determine the location of origins of DNA replication. In the yeast Saccharomyces cerevisiae, the ARS1 replicator contains multiple functional DNA elements: an essential A element and three important B elements--B1, B2, and B3. Functionally similar A, B1, and B2 elements are also present in the ARS307 replicator. The B3 element binds a replication and transcription enhancer protein Abf1p, whereas the A element is required for binding the origin recognition complex (ORC). The function of the B1 and B2 elements remains to be defined. We have used a gel-based DNA binding assay to study the interaction between replicators and the putative initiator protein ORC. In addition to the established requirements for ATP and the A element for ORC-DNA interaction, the new data demonstrate that sequences in the B1 element are also important for ORC-DNA association. This conclusion is supported by DNase I footprint analyses and demonstrates that ORC binds to a bipartitite recognition element within the DNA. Furthermore, mutation of nucleotides in the B1 element suggests that this element has other functions in the initiation of DNA replication besides participating in the ORC-DNA interaction.

Item Type: Paper
Uncontrolled Keywords: Base Sequence Binding Sites Chromosomes Fungal DNA Replication DNA, Fungal metabolism DNA-Binding Proteins biosynthesis metabolism Deoxyribonuclease I Fungal Proteins metabolism Genes Fungal Molecular Sequence Data Oligodeoxyribonucleotides Plasmids Replication Origin Saccharomyces cerevisiae genetics metabolism Saccharomyces cerevisiae Proteins Transcription Factors biosynthesis metabolism
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date: 14 March 1995
Date Deposited: 02 Mar 2012 18:39
Last Modified: 13 Sep 2019 16:43
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