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- W3039200746 abstract "Abstract DNA replication in eukaryotic cells initiates from chromosomal locations, called replication origins, that bind the Origin Recognition Complex (ORC) prior to S phase. Origin establishment is guided by well-defined DNA sequence motifs in Saccharomyces cerevisiae and some other budding yeasts, but most eukaryotes lack sequence-specific origins. At present, the mechanistic and evolutionary reasons for this difference are unclear. A 3.9 Å structure of S. cerevisiae ORC-Cdc6-Cdt1-Mcm2-7 (OCCM) bound to origin DNA revealed, among other things, that a loop within Orc2 inserts into a DNA minor groove and an α-helix within Orc4 inserts into a DNA major groove 1 . We show that this Orc4 α-helix mediates the sequence-specificity of origins in S. cerevisiae . Specifically, mutations were identified within this α-helix that alter the sequence-dependent activity of individual origins as well as change global genomic origin firing patterns. This was accomplished using a massively parallel origin selection assay analyzed using a custom mutual-information-based modeling approach and a separate analysis of whole-genome replication profiling and statistics. Interestingly, the sequence specificity of DNA replication initiation, as mediated by the Orc4 α-helix, has evolved in close conjunction with the gain of ORC-Sir4-mediated gene silencing and the loss of RNA interference." @default.
- W3039200746 created "2020-07-10" @default.
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- W3039200746 date "2020-07-04" @default.
- W3039200746 modified "2023-09-26" @default.
- W3039200746 title "Evolution of DNA Replication Origin Specification and Gene Silencing Mechanisms" @default.
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- W3039200746 doi "https://doi.org/10.1101/2020.07.04.187286" @default.
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