Scientific article
OA Policy
English

The Positioning and Dynamics of Origins of Replication in the Budding Yeast Nucleus

Published inThe Journal of Cell Biology, vol. 152, no. 2, p. 385-400
Publication date2001
Abstract

We have analyzed the subnuclear position of early- and late-firing origins of DNA replication in intact yeast cells using fluorescence in situ hybridization and green fluorescent protein (GFP)-tagged chromosomal domains. In both cases, origin position was determined with respect to the nuclear envelope, as identified by nuclear pore staining or a NUP49-GFP fusion protein. We find that in G1 phase nontelomeric late-firing origins are enriched in a zone immediately adjacent to the nuclear envelope, although this localization does not necessarily persist in S phase. In contrast, early firing origins are randomly localized within the nucleus throughout the cell cycle. If a late-firing telomere-proximal origin is excised from its chromosomal context in G1 phase, it remains late-firing but moves rapidly away from the telomere with which it was associated, suggesting that the positioning of yeast chromosomal domains is highly dynamic. This is confirmed by time-lapse microscopy of GFP-tagged origins in vivo. We propose that sequences flanking late-firing origins help target them to the periphery of the G1-phase nucleus, where a modified chromatin structure can be established. The modified chromatin structure, which would in turn retard origin firing, is both autonomous and mobile within the nucleus.

Keywords
  • Cell cycle
  • Physiology
  • Cell nucleus
  • Genetics
  • Ultrastructure
  • DNA replication
  • G1 phase
  • G2 phase
  • Green fluorescent proteins
  • In situ hybridization
  • Luminescent proteins
  • Analysis
  • Nuclear envelope
  • Recombinant fusion proteins
  • Replication origin
  • S phase
  • Saccharomyces cerevisiae
  • Cytology
Affiliation entities Not a UNIGE publication
Citation (ISO format)
HEUN, Patrick et al. The Positioning and Dynamics of Origins of Replication in the Budding Yeast Nucleus. In: The Journal of Cell Biology, 2001, vol. 152, n° 2, p. 385–400. doi: 10.1083/jcb.152.2.385
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Article (Published version)
accessLevelPublic
Identifiers
Journal ISSN0021-9525
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