Abstract

BackgroundReplication and transcription, the two key functions of DNA, require unwinding of the DNA double helix. It has been shown that replication origins in the budding yeast, Saccharomyces cerevisiae contain an easily unwound stretch of DNA. We have used a recently developed method for determining the locations and degrees of stress-induced duplex destabilization (SIDD) for all the reported replication origins in the genome of the fission yeast, Schizosaccharomyces pombe.ResultsWe have found that the origins are more susceptible to SIDD as compared to the non-origin intergenic regions (NOIRs) and genes. SIDD analysis of many known origins in other eukaryotes suggests that SIDD is a common property of replication origins. Interestingly, the previously shown deletion-dependent changes in the activities of the origins of the ura4 origin region on chromosome 3 are paralleled by changes in SIDD properties, suggesting SIDD’s role in origin activity. SIDD profiling following in silico deletions of some origins suggests that many of the closely spaced S. pombe origins could be clusters of two or three weak origins, similar to the ura4 origin region.ConclusionSIDD appears to be a highly conserved, functionally important property of replication origins in S. pombe and other organisms. The distinctly low SIDD scores of origins and the long range effects of genetic alterations on SIDD properties provide a unique predictive potential to the SIDD analysis. This could be used in exploring different aspects of structural and functional organization of origins including interactions between closely spaced origins.

Highlights

  • Replication and transcription, the two key functions of DNA, require unwinding of the DNA double helix

  • Our results show that S. pombe origins are more susceptible to stress-induced duplex destabilization than their adjacent genes and non-origin intergenes and that, in case of closely spaced origins, the extent of destabilization appears to influence the origin activity

  • stress-induced duplex destabilization (SIDD) analysis of S. cerevisiae replication origins has shown earlier that they are highly susceptible to superhelically driven DNA duplex destabilization [33]

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Summary

Introduction

Replication and transcription, the two key functions of DNA, require unwinding of the DNA double helix. In addition to 54 precisely localized origins by the two-dimensional agarose gel electrophoresis origin mapping technique (2D technique) and DNA combing [18,19,20,21,22,23,24,25,26], different genomewide origin mapping studies have located several hundred to nearly one thousand potential origins in S. pombe [11,25,27,28,29] All these origins are confined to the intergenic regions (IRs), which usually have higher AT content than the genomic average. Replication origins of S. pombe have not been extensively analyzed for their helical stability and the destabilization properties of the origin-containing intergenic regions (OIRs) remain to be known

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