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TitleDNA bending mediated by ORC is essential for replication licensing in budding yeast.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 122, Issue 14, Page e2502277122, Year 2025
Publish dateApr 8, 2025
AuthorsWai Hei Lam / Daqi Yu / Qiongdan Zhang / Yuhan Lin / Ningning Li / Jian Li / Yue Wu / Yingyi Zhang / Ning Gao / Bik Kwoon Tye / Yuanliang Zhai / Shangyu Dang /
PubMed AbstractIn eukaryotes, the origin recognition complex (ORC) promotes the assembly of minichromosome maintenance 2 to 7 complexes into a head-to-head double hexamer at origin DNA in a process known as ...In eukaryotes, the origin recognition complex (ORC) promotes the assembly of minichromosome maintenance 2 to 7 complexes into a head-to-head double hexamer at origin DNA in a process known as replication licensing. In this study, we present a series of cryoelectron microscopy structures of yeast ORC mutants in complex with origin DNA. We show that Orc6, the smallest subunit of ORC, utilizes its transcription factor II B-B domain to orchestrate the sequential binding of ORC to origin DNA. In addition, Orc6 plays the role of a scaffold by stabilizing the basic patch (BP) of Orc5 for ORC to capture and bend origin DNA. Importantly, disrupting DNA bending through mutating three key residues in Orc5-BP impairs ORC's ability to promote replication initiation at two points during the pre-RC assembly process. This study dissects the multifaceted role of Orc6 in orchestrating ORC's activities on DNA and underscores the vital role of DNA bending by ORC in replication licensing.
External linksProc Natl Acad Sci U S A / PubMed:40184174 / PubMed Central
MethodsEM (single particle)
Resolution2.98 - 3.33 Å
Structure data

60326

8zp4

EMDB-60326, PDB-8zp4:
Cryo-EM structure of origin recognition complex (Orc1 to 5) with ARS1 DNA bound
Method: EM (single particle) / Resolution: 3.33 Å

60327

8zp5

EMDB-60327, PDB-8zp5:
Cryo-EM structure of origin recognition complex (Orc5 basic patch mutations) with ARS1 DNA bound
Method: EM (single particle) / Resolution: 2.98 Å

60347

8zpk

EMDB-60347, PDB-8zpk:
Cryo-EM structure of origin recognition complex (Orc6 with residues 1 to 270 deleted) with ARS1 DNA bound
Method: EM (single particle) / Resolution: 3.21 Å

Chemicals

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae (brewer's yeast)
  • saccharomyces cerevisiae s288c (yeast)
KeywordsREPLICATION / origin recognition complex

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