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Molecular and Cellular Biology, July 2007, p. 4652-4663, Vol. 27, No. 13
0270-7306/07/$08.00+0     doi:10.1128/MCB.01246-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Linear Derivatives of Saccharomyces cerevisiae Chromosome III Can Be Maintained in the Absence of Autonomously Replicating Sequence Elements{triangledown} ,{dagger}

Ann Dershowitz,1 Marylynn Snyder,1,{ddagger} Mohammed Sbia,1,§ Joan H. Skurnick,2 Loke Y. Ong,1,¶ and Carol S. Newlon1*

Department of Microbiology and Molecular Genetics,1 Department of Preventive Medicine and Community Health, UMDNJ-New Jersey Medical School, Newark, New Jersey 071032

Received 9 July 2006/ Returned for modification 5 September 2006/ Accepted 16 April 2007

Replication origins in Saccharomyces cerevisiae are spaced at intervals of approximately 40 kb. However, both measurements of replication fork rate and studies of hypomorphic alleles of genes encoding replication initiation proteins suggest the question of whether replication origins are more closely spaced than should be required. We approached this question by systematically deleting replicators from chromosome III. The first significant increase in loss rate detected for the 315-kb full-length chromosome occurred only after all five efficient chromosomal replicators in the left two-thirds of the chromosome (ARS305, ARS306, ARS307, ARS309, and ARS310) had been deleted. The removal of the inefficient replicator ARS308 from this originless region caused little or no additional increase in loss rate. Chromosome fragmentations that removed the normally inactive replicators on the left end of the chromosome or the replicators distal to ARS310 on the right arm showed that both groups of replicators contribute significantly to the maintenance of the originless chromosome. Surprisingly, a 142-kb derivative of chromosome III, lacking all sequences that function as autonomously replicating sequence elements in plasmids, replicated and segregated properly 97% of the time. Both the replication initiation protein ORC and telomeres or a linear topology were required for the maintenance of chromosome fragments lacking replicators.

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, UMDNJ-New Jersey Medical School, ICPH, 225 Warren St., Newark, NJ 07103. Phone: (973) 972-4483, ext. 24227. Fax: (973) 972-1286. E-mail: newlon{at}umdnj.edu

{triangledown} Published ahead of print on 23 April 2007.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} Present address: Department of Physiology and Biophysics, Weill Medical College of Cornell University, 1300 York Ave., New York, NY 10021.

§ Present address: Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, 2000 Circle of Hope, Salt Lake City, UT 84112.

Present address: SciMed Technologies Sdn. Bhd., 1F-08 IOI Business Park, No. 1 Persiaran Puchong Jaya Selatan, Bandar Puchong Jaya, 47100 Puchong, Selangor, Malaysia.

Molecular and Cellular Biology, July 2007, p. 4652-4663, Vol. 27, No. 13
0270-7306/07/$08.00+0     doi:10.1128/MCB.01246-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Di Rienzi, S. C., Collingwood, D., Raghuraman, M. K., Brewer, B. J. (2009). Fragile Genomic Sites Are Associated with Origins of Replication. Gen Biol Evol 2009: 350-363 [Abstract] [Full Text]  
  • Theis, J. F., Dershowitz, A., Irene, C., Maciariello, C., Tobin, M. L., Liberi, G., Tabrizifard, S., Korus, M., Fabiani, L., Newlon, C. S. (2007). Identification of Mutations That Decrease the Stability of a Fragment of Saccharomyces cerevisiae Chromosome III Lacking Efficient Replicators. Genetics 177: 1445-1458 [Abstract] [Full Text]  


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