The requirement of yeast replication origins for pre-replication complex proteins is modulated by transcription

Nieduszynski, Conrad A. ORCID: https://orcid.org/0000-0003-2001-076X, Blow, J. Julian and Donaldson, Anne D. (2005) The requirement of yeast replication origins for pre-replication complex proteins is modulated by transcription. Nucleic Acids Research, 33 (8). pp. 2410-2420. ISSN 0305-1048

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Abstract

The mini-chromosome maintenance proteins Mcm2-7 are essential for DNA replication. They are loaded onto replication origins during G1 phase of the cell cycle to form a pre-replication complex (pre-RC) that licenses each origin for subsequent initiation. We have investigated the DNA elements that determine the dependence of yeast replication origins on Mcm2-7 activity, i.e. the sensitivity of an origin to mcm mutations. Using chimaeric constructs from mcm sensitive and mcm insensitive origins, we have identified two main elements affecting the requirement for Mcm2-7 function. First, transcription into an origin increases its dependence on Mcm2-7 function, revealing a conflict between pre-RC assembly and transcription. Second, sequence elements within the minimal origin influence its mcm sensitivity. Replication origins show similar differences in sensitivity to mutations in other pre-RC proteins (such as Origin Recognition Complex and Cdc6), but not to mutations in initiation and elongation factors, demonstrating that the mcm sensitivity of an origin is determined by its ability to establish a pre-RC. We propose that there is a hierarchy of replication origins with respect to the range of pre-RC protein concentrations under which they will function. This hierarchy is both 'hard-wired' by the minimal origin sequences and 'soft-wired' by local transcriptional context.

Item Type:	Article
Additional Information:	Funding Information: The authors thank Prof. Bik-Kwoon Tye and Dr Tomoyuki Tanaka for strains and plasmids used in this study and Prof. Mike Stark for technical advice. The authors are grateful to Dr John Diffley and Prof. Bik-Kwoon Tye for communicating unpublished results. This work was funded by Cancer Research UK grant C1445/A2570. A.D.D. is a Royal Society University Research Fellow and an EMBO Young Investigator. J.J.B. is funded by Cancer Research UK grants SP2517/0101 and C303/A3135. Funding to pay the Open Access publication charges for this article was provided by JISC.
Uncontrolled Keywords:	genetics ,/dk/atira/pure/subjectarea/asjc/1300/1311
Faculty \ School:	Faculty of Science > School of Biological Sciences
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	08 Sep 2022 08:31
Last Modified:	21 Oct 2022 01:37
URI:	https://ueaeprints.uea.ac.uk/id/eprint/87896
DOI:	10.1093/nar/gki539

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