Specific basic patch-dependent multimerization of Saccharomyces cerevisiae ORC on single-stranded DNA promotes ATP hydrolysis.

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ID: 45330
2019
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Abstract
Replication initiation at specific genomic loci dictates precise duplication and inheritance of genetic information. In eukaryotic cells, ATP-bound origin recognition complexes (ORCs) stably bind to double-stranded (ds) DNA origins to recruit the replicative helicase onto the origin DNA. To achieve these processes, an essential region of the origin DNA must be recognized by the eukaryotic origin sensor (EOS) basic patch within the disordered domain of the largest ORC subunit, Orc1. Although ORC also binds single-stranded (ss) DNA in an EOS-independent manner, it is unknown whether EOS regulates ORC on ssDNA. We found that, in budding yeast, ORC multimerizes on ssDNA in vitro independently of adenine nucleotides. We also found that the ORC multimers form in an EOS-dependent manner and stimulate the ORC ATPase activity. An analysis of genomics data supported the idea that ORC-ssDNA binding occurs in vivo at specific genomic loci outside of replication origins. These results suggest that EOS function is differentiated by ORC-bound ssDNA, which promotes ORC self-assembly and ATP hydrolysis. These mechanisms could modulate ORC activity at specific genomic loci and could be conserved among eukaryotes.
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kawakami2019specificgenes Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Kawakami, Hironori;Muraoka, Ryuya;Ohashi, Eiji;Kawabata, Kenta;Kanamoto, Shota;Chichibu, Takeaki;Tsurimoto, Toshiki;Katayama, Tsutomu;
Journal genes to cells : devoted to molecular & cellular mechanisms
Year 2019
DOI
10.1111/gtc.12710
URL
Keywords
intrinsically disordered region aaa+ atpase dna replication origin recognition complex protein multimerization single-stranded dna

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