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Related Experiment Videos

The origin recognition complex in silencing, cell cycle progression, and DNA replication

S Loo1, C A Fox, J Rine

  • 1Department of Molecular and Cell Biology, University of California, Berkeley 94720, USA.

Molecular Biology of the Cell
|June 1, 1995
PubMed
Summary

The origin recognition complex subunit 5 (ORC5) is crucial for DNA replication and cell cycle progression. Mutations in ORC5 disrupt these processes and affect gene silencing.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • The origin recognition complex (ORC) is essential for initiating DNA replication in eukaryotes.
  • ORC is a multi-subunit complex, but the specific roles of individual subunits are not fully understood.
  • Previous studies suggested ORC involvement in DNA replication and gene silencing.

Purpose of the Study:

  • To isolate and characterize the ORC5 gene and its corresponding protein subunit.
  • To investigate the function of ORC5 in DNA replication and cell cycle control.
  • To explore the role of ORC5 in gene silencing.

Main Methods:

  • Isolation and characterization of ORC5 gene mutants (orc5-1).
  • Phenotypic analysis of orc5-1 mutants, including growth, cell cycle progression, and plasmid stability.

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  • Genetic interaction studies with other cell cycle genes.
  • Sequence analysis of ORC5 to identify functional domains.

    • Main Results:

    • The orc5-1 mutation confers temperature-sensitive growth and cell cycle arrest at restrictive temperatures.
    • orc5-1 mutants exhibit increased plasmid loss, suppressible by additional replication origins.
    • ORC5 sequence analysis revealed a putative ATP-binding site, suggesting a role in ATP-dependent ORC-origin binding.
    • Genetic interactions and silencing defects link ORC5 and the ORC complex to DNA replication and silencing.

    Conclusions:

    • ORC5 is a vital subunit of the origin recognition complex, essential for DNA replication initiation.
    • The ORC complex, including ORC5, plays a dual role in both DNA replication and gene silencing.
    • Orc5p's potential ATP-binding site suggests a mechanism for ORC's origin association.