The fission yeast homologue of Orc4p binds to replication origin DNA via multiple AT-hooks

Chuang, Ronald Y.; Kelly, Thomas J.

doi:10.1073/pnas.96.6.2656

Cited by 223 publications

(167 citation statements)

References 55 publications

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“…In many cases, especially ORIs that map in large IGRs, they are made up of a variable number of independent A þT-rich modules that can recruit ORC through the Orc4 subunit whose 9 AT-hook domains are sufficient to target ORC to A þT-rich sequences (Chuang and Kelly, 1999;Lee et al, 2001). The simplest possibility to explain the biased localization of ORIs is that, given that the average A þT content of IGRs is 70%, large IGRs will have a higher probability of including favourable combinations of A þT-rich elements capable of recruiting ORC.…”

Section: Discussionmentioning

confidence: 99%

Nucleosomal organization of replication origins and meiotic recombination hotspots in fission yeast

et al. 2011

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In Schizosaccharomyces pombe, DNA replication origins (ORIs) and meiotic recombination hotspots lack consensus sequences and show a bias towards mapping to large intergenic regions (IGRs). To explore whether this preference depended on underlying chromatin features, we have generated genome-wide nucleosome profiles during mitosis and meiosis. We have found that meiotic double-strand break sites (DSBs) colocalize with nucleosome-depleted regions (NDRs) and that large IGRs include clusters of NDRs that overlap with almost half of all DSBs. By contrast, ORIs do not colocalize with NDRs and they are regulated independently of DSBs. Physical relocation of NDRs at ectopic loci or modification of their genomic distribution during meiosis was paralleled by the generation of new DSB sites. Over 80% of all meiotic DSBs colocalize with NDRs that are also present during mitosis, indicating that the recombination pattern is largely dependent on constitutive properties of the genome and, to a lesser extent, on the transcriptional profile during meiosis. The organization of ORIs and of DSBs regions in S. pombe reveals similarities and differences relative to Saccharomyces cerevisiae.

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Section: Discussionmentioning

confidence: 99%

Nucleosomal organization of replication origins and meiotic recombination hotspots in fission yeast

et al. 2011

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“…The binding through the GAL4 DNA-binding site still allows replication initiation. The GAL4 DNA-binding domain seems analogous to the naturally occurring AT-hook domain at the N terminus of Orc4 in Schizosaccharomyces pombe that has been shown to recruit ORC to AT-rich DNA segments that specify origins in S. pombe (Chuang and Kelly 1999;Lee et al 2001). Orc4 from other species lacks this AThook domain.…”

Section: An Artificial Replication Origin Created In Vivo Genes and Devmentioning

confidence: 99%

Recruitment of ORC or CDC6 to DNA is sufficient to create an artificial origin of replication in mammalian cells

Takeda¹,

Shibata²,

Parvin³

et al. 2005

Genes Dev.

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Origins of replication are expected to recruit initiation proteins like origin recognition complex (ORC) and Cdc6 in eukaryotes and provide a platform for unwinding DNA. Here we test whether localization of initiation proteins onto DNA is sufficient for origin function. Different components of the ORC complex and Cdc6 stimulated prereplicative complex (pre-RC) formation and replication initiation when fused to the GAL4 DNA-binding domain and recruited to plasmid DNA containing a tandem array of GAL4-binding sites. Replication occurred once per cell cycle and was inhibited by Geminin, indicating that the plasmid was properly licensed during the cell cycle. The GAL4 fusion protein recruits other polypeptides of the ORC-Cdc6 complex, and nascent strand abundance was highest near the GAL4-binding sites. Therefore, the artificial origin recapitulates many of the regulatory features of physiological origins and is valuable for studies on replication initiation in mammalian cells. We demonstrated the utility of this system by showing the functional importance of the ATPase domains of human Cdc6 and Orc1 and the dispensability of the N-terminal segments of Orc1 and Orc2 in this assay. Artificial recruitment of a eukaryotic cellular replication initiation factor to a DNA sequence can create a functional origin of replication, providing a robust genetic assay for these factors and a novel approach to generating episomal vectors for gene therapy. In order to duplicate the massive eukaryotic genome in a reasonable amount of time, DNA replication begins from multiple sites located throughout the genome called replication origins (for review, see Gilbert 2001). In the budding yeast, Saccharomyces cerevisiae, replication origins are defined by specific DNA sequences ∼100 base pairs (bp) in length. Plasmids containing these sequences replicate when transformed into cells and hence are referred to as autonomously replicating sequences, or ARS. Within each ARS is an 11-bp ARS consensus sequence (ACS) that is required for binding of the six-subunit origin recognition complex (ORC). ORC remains bound to chromatin throughout the cell cycle in S. cerevisiae, where it catalyzes the ordered assembly of replication factors that culminate in the loading of the replicative polymerases that begin DNA synthesis (Bell and Dutta 2002;Mendez and Stillman 2003). Although ORC and many of the proteins involved in replication are conserved in all eukaryotes, identification of conserved DNA sequences that define eukaryotic origins remains elusive (Gilbert 2004;Cvetic and Walter 2005). Similar plasmid transformation experiments that were successful in budding yeast have failed to identify ARS sequences in other eukaryotes. In mammalian cells, replication initiation occurs from poorly localized replication zones of 6-55 kb, although in a few cases the origin has been mapped to smaller lengths of DNA. Having welldefined origins in S. cerevisiae has greatly facilitated studying replication initiation. The initial discovery and characterization of ORC w...

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“…In S. cerevisiae, ORC can bind sequence specifically to the consensus sequence within autonomously replicating sequences (3,8). In another yeast, Schizosaccharomyces pombe, ORC can recognize the AT-tracts present in autonomously replicating sequences; the AT-hook motif of the ORC4 subunit is responsible for this recognition (9).…”

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confidence: 99%

Human Origin Recognition Complex Binds Preferentially to G-quadruplex-preferable RNA and Single-stranded DNA

Hoshina

Yura

Teranishi

et al. 2013

Journal of Biological Chemistry

103

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Background: ORC binds to replication origins, but human ORC does not exhibit apparent sequence-specificity. Results: G-quadruplex (G4)-preferable RNA or single-stranded DNA competes for DNA binding of ORC. Conclusion: Human ORC binds preferentially to RNA and single-stranded DNA that form G4, and the certain domain in ORC1 is involved in this binding. Significance: This ability may correlate with the G4-formable motif in human replication origins.

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The fission yeast homologue of Orc4p binds to replication origin DNA via multiple AT-hooks

Abstract: The origin recognition complex (ORC) was originally identified in the yeast Saccharomyces cerevisiae as a protein that specifically binds to origins of DNA replication.

Cited by 223 publications

References 55 publications

Nucleosomal organization of replication origins and meiotic recombination hotspots in fission yeast

Nucleosomal organization of replication origins and meiotic recombination hotspots in fission yeast

Recruitment of ORC or CDC6 to DNA is sufficient to create an artificial origin of replication in mammalian cells

Human Origin Recognition Complex Binds Preferentially to G-quadruplex-preferable RNA and Single-stranded DNA

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