The effects of manipulating levels of replication initiation factors on origin firing efficiency in yeast

English version

Autoři: Kelsey L. Lynch ^aff001; Gina M. Alvino ^aff002; Elizabeth X. Kwan ^aff002; Bonita J. Brewer ^aff001; M. K. Raghuraman ^aff002
Působiště autorů: Molecular and Cellular Biology Program, University of Washington, Seattle, Washington, United States of America ^aff001; Department of Genome Sciences, University of Washington, Seattle, Washington, United States of America ^aff002
Vyšlo v časopise: The effects of manipulating levels of replication initiation factors on origin firing efficiency in yeast. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008430
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1008430

Souhrn

Chromosome replication in Saccharomyces cerevisiae is initiated from ~300 origins that are regulated by DNA sequence and by the limited abundance of six trans-acting initiation proteins (Sld2, Sld3, Dpb11, Dbf4, Sld7 and Cdc45). We set out to determine how the levels of individual factors contribute to time of origin activation and/or origin efficiency using induced depletion of single factors and overexpression of sets of multiple factors. Depletion of Sld2 or Sld3 slows growth and S phase progression, decreases origin efficiency across the genome and impairs viability as a result of incomplete replication of the rDNA. We find that the most efficient early origins are relatively unaffected by depletion of either Sld2 or Sld3. However, Sld3 levels, and to a lesser extent Sld2 levels, are critical for firing of the less efficient early origins. Overexpression of Sld3 simultaneously with Sld2, Dpb11 and Dbf4 preserves the relative efficiency of origins. Only when Cdc45 and Sld7 are also overexpressed is origin efficiency equalized between early- and late-firing origins. Our data support a model in which Sld3 together with Cdc45 (and/or Sld7) is responsible for the differential efficiencies of origins across the yeast genome.

Klíčová slova:

DNA replication – Saccharomyces cerevisiae – Cell cycle and cell division – Hyperexpression techniques – Flow cytometry – Gel electrophoresis – Auxins – Synthesis phase

Zdroje

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