Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo.

2.50
Hdl Handle:
http://hdl.handle.net/10541/82357
Title:
Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo.
Authors:
Kanemaki, Masato; Sanchez-Diaz, Alberto; Gambus, Agnieszka; Labib, Karim
Abstract:
Evolutionarily diverse eukaryotic cells share many conserved proteins of unknown function. Some are essential for cell viability, emphasising their importance for fundamental processes of cell biology but complicating their analysis. We have developed an approach to the large-scale characterization of such proteins, based on conditional and rapid degradation of the target protein in vivo, so that the immediate consequences of bulk protein depletion can be examined. Budding yeast strains have been constructed in which essential proteins of unknown function have been fused to a 'heat-inducible-degron' cassette that targets the protein for proteolysis at 37 degrees C (ref. 4). By screening the collection for defects in cell-cycle progression, here we identify three DNA replication factors that interact with each other and that have uncharacterized homologues in human cells. We have used the degron strains to show that these proteins are required for the establishment and normal progression of DNA replication forks. The degron collection could also be used to identify other, essential, proteins with roles in many other processes of eukaryotic cell biology.
Affiliation:
Cancer Research UK, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester, M20 4BX, UK.
Citation:
Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo. 2003, 423 (6941):720-4 Nature
Journal:
Nature
Issue Date:
12-Jun-2003
URI:
http://hdl.handle.net/10541/82357
DOI:
10.1038/nature01692
PubMed ID:
12768207
Type:
Article
Language:
en
ISSN:
0028-0836
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorKanemaki, Masato-
dc.contributor.authorSanchez-Diaz, Alberto-
dc.contributor.authorGambus, Agnieszka-
dc.contributor.authorLabib, Karim-
dc.date.accessioned2009-09-23T13:27:46Z-
dc.date.available2009-09-23T13:27:46Z-
dc.date.issued2003-06-12-
dc.identifier.citationFunctional proteomic identification of DNA replication proteins by induced proteolysis in vivo. 2003, 423 (6941):720-4 Natureen
dc.identifier.issn0028-0836-
dc.identifier.pmid12768207-
dc.identifier.doi10.1038/nature01692-
dc.identifier.urihttp://hdl.handle.net/10541/82357-
dc.description.abstractEvolutionarily diverse eukaryotic cells share many conserved proteins of unknown function. Some are essential for cell viability, emphasising their importance for fundamental processes of cell biology but complicating their analysis. We have developed an approach to the large-scale characterization of such proteins, based on conditional and rapid degradation of the target protein in vivo, so that the immediate consequences of bulk protein depletion can be examined. Budding yeast strains have been constructed in which essential proteins of unknown function have been fused to a 'heat-inducible-degron' cassette that targets the protein for proteolysis at 37 degrees C (ref. 4). By screening the collection for defects in cell-cycle progression, here we identify three DNA replication factors that interact with each other and that have uncharacterized homologues in human cells. We have used the degron strains to show that these proteins are required for the establishment and normal progression of DNA replication forks. The degron collection could also be used to identify other, essential, proteins with roles in many other processes of eukaryotic cell biology.en
dc.language.isoenen
dc.subject.meshCell Cycle Proteins-
dc.subject.meshCell Survival-
dc.subject.meshDNA Replication-
dc.subject.meshDoxycycline-
dc.subject.meshEndopeptidases-
dc.subject.meshGene Expression Regulation-
dc.subject.meshGenes, Essential-
dc.subject.meshLigases-
dc.subject.meshProtein Processing, Post-Translational-
dc.subject.meshProteome-
dc.subject.meshProteomics-
dc.subject.meshReplication Origin-
dc.subject.meshSaccharomyces cerevisiae-
dc.subject.meshSaccharomyces cerevisiae Proteins-
dc.subject.meshTemperature-
dc.subject.meshUbiquitin-Protein Ligases-
dc.titleFunctional proteomic identification of DNA replication proteins by induced proteolysis in vivo.en
dc.typeArticleen
dc.contributor.departmentCancer Research UK, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester, M20 4BX, UK.en
dc.identifier.journalNatureen

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