Ease of DNA unwinding is a conserved property of yeast replication origins (original) (raw)

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Molecular and Cellular Biology Department, Roswell Park Cancer lnstftute

Buffalo, NY 14263, USA

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Molecular and Cellular Biology Department, Roswell Park Cancer lnstftute

Buffalo, NY 14263, USA

• Present address: Biology Department, University of Virginia, Charlottesville VA 22903, USA

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Molecular and Cellular Biology Department, Roswell Park Cancer lnstftute

Buffalo, NY 14263, USA

* To whom correspondence should be addressed

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• Present address: Biology Department, University of Virginia, Charlottesville VA 22903, USA

Author Notes

Received:

16 October 1992

Revision received:

18 December 1992

Accepted:

18 December 1992

Published:

11 February 1993

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Darren A. Natale, Robert M. Umek, David Kowalski, Ease of DNA unwinding is a conserved property of yeast replication origins, Nucleic Acids Research, Volume 21, Issue 3, 11 February 1993, Pages 555–560, https://doi.org/10.1093/nar/21.3.555
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Abstract

Autonomously replicating sequence ( ARS ) elements function as plasmid replication origins. Our studies of the H4 ARS and ARS3O7 have established the requirement for a DNA unwinding element (DUE), a broad easily-unwound sequence 3′ to the essential consensus that likely facilitates opening of the origin. in this report, we examine the intrinsic ease of unwinding a variety of ARS elements using (1) a single-strand-specific nuclease to probe for DNA unwinding in a negatively-supercoiled piasmid, and (2) a computer program that calculates DNA helical stability from the nucleotide sequence. ARS elements that are associated with replication origins on chromosome III are nuclease hypersensitive, and the helical stability minima correctly predict the location and hierarchy of the hypersensitive sites. All well-studied ARS elements in which the essential consensus sequence has been identified by mutational analysis contain a 100-bp region of low helical stability immediately 3′ to the consensus, as do ARS elements created by mutation within the prokaryotic M13 vector. The level of helical stability is, in all cases, below that of ARS3O7 derivatives inactivated by mutations in the DUE. Our findings indicate that the ease of DNA unwinding at the broad region directly 3′ to the ARS consensus is a conserved property of yeast replication origins.

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Author notes

• Present address: Biology Department, University of Virginia, Charlottesville VA 22903, USA

© 1993 Oxford University Press

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