Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo.
AffiliationCancer Research UK, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester, M20 4BX, UK.
MetadataShow full item record
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.
CitationFunctional proteomic identification of DNA replication proteins by induced proteolysis in vivo. 2003, 423 (6941):720-4 Nature
- Rapid depletion of budding yeast proteins by fusion to a heat-inducible degron.
- Authors: Sanchez-Diaz A, Kanemaki M, Marchesi V, Labib K
- Issue date: 2004 Mar 2
- Cyclin-dependent kinase and Cks/Suc1 interact with the proteasome in yeast to control proteolysis of M-phase targets.
- Authors: Kaiser P, Moncollin V, Clarke DJ, Watson MH, Bertolaet BL, Reed SI, Bailly E
- Issue date: 1999 May 1
- Uninterrupted MCM2-7 function required for DNA replication fork progression.
- Authors: Labib K, Tercero JA, Diffley JF
- Issue date: 2000 Jun 2
- Mcm10 plays a role in functioning of the eukaryotic replicative DNA helicase, Cdc45-Mcm-GINS.
- Authors: Watase G, Takisawa H, Kanemaki MT
- Issue date: 2012 Feb 21
- The anaphase-promoting complex is required in G1 arrested yeast cells to inhibit B-type cyclin accumulation and to prevent uncontrolled entry into S-phase.
- Authors: Irniger S, Nasmyth K
- Issue date: 1997 Jul