Doctoral thesis
Open access

Spatiotemporal characterization of DNA replication in the C. elegans embryo

ContributorsStrobino, Maude
Defense date2020-04-01

Histone H3.3 is a variant of histone H3. Here we show that loss of H3.3 results in replication defects in Caenorhabditis elegans embryos at elevated temperatures. To characterize these defects, we adapt methods to determine replication timing, map replication origins, and examine replication fork progression. Our analysis shows that the genome is replicated from early and late firing origins and is partitioned into domains of early and late replication. We find that under temperature stress conditions, additional replication origins become activated. Moreover, loss of H3.3 results in altered replication fork progression around origins. These replication defects are accompanied by replication checkpoint activation, a delayed cell cycle, and increased lethality in checkpoint-compromised embryos. Our comprehensive analysis of DNA replication in C. elegans reveals the genomic location of replication origins and the dynamics of their firing, and uncovers a role of H3.3 in the regulation of replication origins under stress conditions.

  • Histone H3.3
  • Replication
  • Origins
  • Repli-seq
  • EdU-seq
  • ChEC-seq
  • DNA combing
  • C. elegans
Citation (ISO format)
STROBINO, Maude. Spatiotemporal characterization of DNA replication in the C. elegans embryo. 2020. doi: 10.13097/archive-ouverte/unige:140702
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Creation08/24/2020 3:32:00 PM
First validation08/24/2020 3:32:00 PM
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