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

Hironori Kawakami, Ryuya Muraoka, Eiji Ohashi, Kenta Kawabata, Shota Kanamoto, Takeaki Chichibu, Toshiki Tsurimoto, Tsutomu Katayama

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)608-618
Number of pages11
JournalGenes to Cells
Volume24
Issue number9
DOIs
Publication statusPublished - Sep 1 2019

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Origin Recognition Complex
Single-Stranded DNA
Saccharomyces cerevisiae
Hydrolysis
Adenosine Triphosphate
DNA
Saccharomycetales
Replication Origin
Adenine Nucleotides
Eukaryotic Cells
Genomics
Eukaryota
Adenosine Triphosphatases

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

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Specific basic patch-dependent multimerization of Saccharomyces cerevisiae ORC on single-stranded DNA promotes ATP hydrolysis. / Kawakami, Hironori; Muraoka, Ryuya; Ohashi, Eiji; Kawabata, Kenta; Kanamoto, Shota; Chichibu, Takeaki; Tsurimoto, Toshiki; Katayama, Tsutomu.

In: Genes to Cells, Vol. 24, No. 9, 01.09.2019, p. 608-618.

Research output: Contribution to journalArticle

Kawakami, Hironori ; Muraoka, Ryuya ; Ohashi, Eiji ; Kawabata, Kenta ; Kanamoto, Shota ; Chichibu, Takeaki ; Tsurimoto, Toshiki ; Katayama, Tsutomu. / Specific basic patch-dependent multimerization of Saccharomyces cerevisiae ORC on single-stranded DNA promotes ATP hydrolysis. In: Genes to Cells. 2019 ; Vol. 24, No. 9. pp. 608-618.
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