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PMID: 17145765 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

The Caenorhabditis elegans replication licensing factor CDT-1 is targeted for degradation by the CUL-4/DDB-1 complex.

Molecular and cellular biology ·Vol. 27 ·No. 4 ·2007-02-00 ·Pages 1394-406

Kim Y, Kipreos ET

Abstract

The replication of genomic DNA is strictly regulated to occur only once per cell cycle. This regulation centers on the temporal restriction of replication licensing factor activity. Two distinct ubiquitin ligase (E3) complexes, CUL4/DDB1 and SCF(Skp2), have been reported to target the replication licensing factor Cdt1 for ubiquitin-mediated proteolysis. However, it is unclear to what extent these two distinct Cdt1 degradation pathways are conserved. Here, we show that Caenorhabditis elegans DDB-1 is required for the degradation of CDT-1 during S phase. DDB-1 interacts specifically with CUL-4 but not with other C. elegans cullins. A ddb-1 null mutant exhibits extensive DNA rereplication in postembryonic BLAST cells, similar to what is observed in cul-4(RNAi) larvae. DDB-1 physically associates with CDT-1, suggesting that CDT-1 is a direct substrate of the CUL-4/DDB-1 E3 complex. In contrast, a deletion mutant of the C. elegans Skp2 ortholog, skpt-1, appears overtly wild type with the exception of an impenetrant gonad migration defect. There is no appreciable role for SKPT-1 in the degradation of CDT-1 during S phase, even in a sensitized ddb-1 mutant background. We propose that the CUL-4/DDB-1 ubiquitin ligase is the principal E3 for regulating the extent of DNA replication in C. elegans.

MeSH Terms
Alleles Animals Caenorhabditis elegans/cytology,metabolism Caenorhabditis elegans Proteins/chemistry,metabolism Cyclin-Dependent Kinase Inhibitor Proteins/metabolism Female Gene Deletion Gene Expression Regulation, Developmental Germ Cells/growth & development Humans Larva/cytology Ligases/chemistry,metabolism Phenotype Protein Binding Protein Processing, Post-Translational S Phase S-Phase Kinase-Associated Proteins/metabolism Vulva/cytology,growth & development
Chemicals
Caenorhabditis elegans Proteins Cdt-1 protein, C elegans Cul-4 protein, C elegans Cyclin-Dependent Kinase Inhibitor Proteins DDB-1 protein, C elegans S-Phase Kinase-Associated Proteins cki-1 protein, C elegans Ligases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kim Youngjo
Department of Cellular Biology, University of Georgia, Athens, GA 30602-2607, USA.
Kipreos Edward T
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.00736-06
Published
2007-02-00
Epub
2006-00-04
Pages
1394-406
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1800708
Subset
IM
Grants
NIGMS NIH HHS · R01 GM055297 · United States
NIGMS NIH HHS · R01 GM 055297 · United States
External Links
PubMed source DOI: 10.1128/MCB.00736-06 PMC Full Text View journal details
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