Dataset Information

CRL4^DCAF1 ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

ABSTRACT: Centrioles play important roles in the assembly of centrosomes and cilia. Centriole duplication occurs once per cell cycle and is dependent on polo-like kinase 4 (PLK4). To prevent centriole amplification, which is a hallmark of cancer, PLK4 protein levels need to be tightly regulated. Here, we show that the Cullin4A/B-DDB1-DCAF1, CRL4^DCAF1, E3 ligase targets PLK4 for degradation in human cells. DCAF1 binds and ubiquitylates PLK4 in the G2 phase to prevent premature centriole duplication in mitosis. In contrast to the regulation of PLK4 by SCF^β-TrCP, the interaction between PLK4 and DCAF1 is independent of PLK4 kinase activity and mediated by polo-boxes 1 and 2 of PLK4, suggesting that DCAF1 promotes PLK4 ubiquitylation independently of β-TrCP. Thus, the SCF^{Slimb/β-TrCP} pathway, targeting PLK4 for ubiquitylation based on its phosphorylation state and CRL4^DCAF1, which ubiquitylates PLK4 by binding to the conserved PB1-PB2 domain, appear to be complementary ways to control PLK4 abundance to prevent centriole overduplication.

SUBMITTER: Grossmann J

PROVIDER: S-EPMC10942865 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature

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CRL4<sup>DCAF1</sup> ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

Grossmann Josina J Kratz Anne-Sophie AS Kordonsky Alina A Prag Gali G Hoffmann Ingrid I

Life science alliance 20240315 6

Centrioles play important roles in the assembly of centrosomes and cilia. Centriole duplication occurs once per cell cycle and is dependent on polo-like kinase 4 (PLK4). To prevent centriole amplification, which is a hallmark of cancer, PLK4 protein levels need to be tightly regulated. Here, we show that the Cullin4A/B-DDB1-DCAF1, CRL4<sup>DCAF1</sup>, E3 ligase targets PLK4 for degradation in human cells. DCAF1 binds and ubiquitylates PLK4 in the G2 phase to prevent premature centriole duplicat ...[more]

PMID: 38490717

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Project description:BackgroundHIV-1 Vpr-mediated G2 cell cycle arrest is dependent on the interaction of Vpr with an E3 ubiquitin ligase that contains damage-specific DNA binding protein 1 (DDB1), Cullin 4A (Cul4A), DDB1 and Cul4-associated factor 1 (DCAF1), and Rbx1. Vpr is thought to associate directly with DCAF1 in the E3 ubiquitin ligase complex although the exact interaction pattern of the proteins in the complex is not completely defined. The Vpr of SIVagm induces G2 arrest of cognate African Green Monkey (AGM) cells but not human cells. The molecular mechanism by which SIVagm Vpr exhibits its species-specific function remained unknown.MethodsPhysical interaction of proteins in the E3 ubiquitin ligase complex was assessed by co-immunoprecipitation followed by western blotting. In addition, co-localization of the proteins in cells was investigated by confocal microscopy. The cell cycle was analyzed by propidium iodide staining and flow cytometry. DNA damage response elicited by Vpr was evaluated by detecting phosphorylation of H2AX, a marker for DNA damage response.ResultsWe show that RNAi knock-down of DCAF1 prevented the co-immunoprecipitation of DDB1 with HIV-1 Vpr while DDB1 knock-down did not influence the binding of Vpr to DCAF1. HIV-1 Vpr mutants with a L64P or a R90K mutation maintained the ability to associate with DCAF1 but did not appear to be in a complex with DDB1. SIVagm Vpr associated with AGM DCAF1 and DDB1 while, in human cells, it binds to human DCAF1 but hardly binds to human DDB1, resulting in the reduced activation of H2AX.ConclusionsThe identification of Vpr mutants which associate with DCAF1 but only poorly with DDB1 suggests that DCAF1 is necessary but the simple binding of Vpr to DCAF1 is not sufficient for the Vpr association with DDB1-containing E3 ligase complex. Vpr may interact both with DCAF1 and DDB1 in the E3 ligase complex. Alternatively, the interaction of Vpr and DCAF1 may induce a conformational change in DCAF1 or Vpr that promotes the interaction with DDB1. The ability of SIVagm Vpr to associate with DDB1, but not DCAF1, can explain the species-specificity of SIVagm Vpr-mediated G2 arrest.

Dataset Information

CRL4^DCAF1 ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

Publications

CRL4<sup>DCAF1</sup> ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

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Dataset Information

CRL4DCAF1 ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

Publications

CRL4<sup>DCAF1</sup> ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

CRL4^DCAF1 ubiquitin ligase regulates PLK4 protein levels to prevent premature centriole duplication.