CUL-4 ubiquitin ligase maintains genome stability by restraining DNA-replication licensing.
To maintain genome stability, DNA replication is strictly regulated to occur only once per cell cycle. In eukaryotes, the presence of 'licensing proteins' at replication origins during the G1 cell-cycle phase allows the formation of the pre-replicative complex. The removal of licensing proteins from chromatin during the S phase ensures that origins fire only once per cell cycle. Here we show that the CUL-4 ubiquitin ligase temporally restricts DNA-replication licensing in Caenorhabditis elegans. Inactivation of CUL-4 causes massive DNA re-replication, producing cells with up to 100C DNA content. The C. elegans orthologue of the replication-licensing factor Cdt1 (refs 2, 3) is required for DNA replication. C. elegans CDT-1 is present in G1-phase nuclei but disappears as cells enter S phase. In cells lacking CUL-4, CDT-1 levels fail to decrease during S phase and instead remain constant in the re-replicating cells. Removal of one genomic copy of cdt-1 suppresses the cul-4 re-replication phenotype. We propose that CUL-4 prevents aberrant re-initiation of DNA replication, at least in part, by facilitating the degradation of CDT-1.
Zhong
Weiwei
W
Department of Cellular Biology, University of Georgia, Athens, Georgia 30602, USA.
Feng
Hui
H
Santiago
Fernando E
FE
Kipreos
Edward T
ET
D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D004261:DNA Replication; D018509:DNA, Helminth / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D016678:Genome; D008025:Ligases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D018510:RNA, Helminth / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases
D029742:Caenorhabditis elegans Proteins; C475683:Cdt-1 protein, C elegans; C475682:Cul-4 protein, C elegans; D018509:DNA, Helminth; D018510:RNA, Helminth; D012333:RNA, Messenger; D044767:Ubiquitin-Protein Ligases; D008025:Ligases