NDLI: DNA helical stability accounts for mutational defects in a yeast replication origin.

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DNA helical stability accounts for mutational defects in a yeast replication origin.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Natale, D. A. Schubert, A. E. Kowalski, D.
Description	Author Affiliation: Natale DA ( Molecular and Cellular Biology Department, Roswell Park Cancer Institute, Buffalo, NY 14263.);
Abstract	Earlier studies on the H4 autonomously replicating sequence (ARS) identified a DNA unwinding element (DUE), a required sequence that is hypersensitive to single-strand-specific nucleases and serves to facilitate origin unwinding. Here we demonstrate that a DUE can be identified in the C2G1 ARS, a chromosomal replication origin, by using a computer program that calculates DNA helical stability from the base sequence. The helical stability minima correctly predict the location and hierarchy of the nuclease-hypersensitive sites in a C2G1 ARS plasmid. Nucleotide-level mapping shows that the nuclease-hypersensitive site at the ARS spans a 100-base-pair sequence in the required 3'-flanking region. Mutations that stabilize the DNA helix in the broad 3'-flanking region reduce or abolish ARS-mediated plasmid replication, indicating that helical instability is required for origin function. The level of helical instability is quantitatively related to the replication efficiency of the ARS mutants. Multiple copies of either a consensus-related sequence present in the C2G1 ARS or the consensus sequence itself in synthetic ARS elements contribute to DNA helical instability. Our findings indicate that a DUE is a conserved component of the C2G1 ARS and is a major determinant of replication origin activity.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	7
Volume Number	89
Language	English
Publisher	National Academy of Sciences
Publisher Date	1992-05-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA Replication DNA, Fungal Chemistry Nucleic Acid Conformation Regulatory Sequences, Nucleic Acid Saccharomyces Cerevisiae Genetics Endodeoxyribonucleases Pharmacology Molecular Sequence Data Structure-Activity Relationship Thermodynamics Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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