RRC ID 34812
Author Kawakami H, Ohashi E, Kanamoto S, Tsurimoto T, Katayama T.
Title Specific binding of eukaryotic ORC to DNA replication origins depends on highly conserved basic residues.
Journal Sci Rep
Abstract In eukaryotes, the origin recognition complex (ORC) heterohexamer preferentially binds replication origins to trigger initiation of DNA replication. Crystallographic studies using eubacterial and archaeal ORC orthologs suggested that eukaryotic ORC may bind to origin DNA via putative winged-helix DNA-binding domains and AAA+ ATPase domains. However, the mechanisms how eukaryotic ORC recognizes origin DNA remain elusive. Here, we show in budding yeast that Lys-362 and Arg-367 residues of the largest subunit (Orc1), both outside the aforementioned domains, are crucial for specific binding of ORC to origin DNA. These basic residues, which reside in a putative disordered domain, were dispensable for interaction with ATP and non-specific DNA sequences, suggesting a specific role in recognition. Consistent with this, both residues were required for origin binding of Orc1 in vivo. A truncated Orc1 polypeptide containing these residues solely recognizes ARS sequence with low affinity and Arg-367 residue stimulates sequence specific binding mode of the polypeptide. Lys-362 and Arg-367 residues of Orc1 are highly conserved among eukaryotic ORCs, but not in eubacterial and archaeal orthologs, suggesting a eukaryote-specific mechanism underlying recognition of replication origins by ORC.
Volume 5
Pages 14929
Published 2015-10-12
DOI 10.1038/srep14929
PII srep14929
PMID 26456755
PMC PMC4601075
MeSH Amino Acid Sequence Arginine / chemistry Arginine / metabolism Conserved Sequence DNA / chemistry* DNA / genetics DNA / metabolism DNA Replication* Lysine / chemistry Lysine / metabolism Models, Molecular Molecular Sequence Data Origin Recognition Complex / chemistry* Origin Recognition Complex / genetics Origin Recognition Complex / metabolism Protein Binding Replication Origin* Saccharomyces cerevisiae / genetics* Saccharomyces cerevisiae / metabolism Saccharomyces cerevisiae Proteins / chemistry* Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism
IF 4.122
Times Cited 2
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Yeast BY5589