CRL4WDR1 Controls Polo-like Kinase Protein Abundance to Promote Bilobe Duplication, Basal Body Segregation and Flagellum Attachment in Trypanosoma brucei

Hu, Huiqing; Zhou, Qing; Han, Xianxian; Li, Ziyin

doi:10.1371/journal.ppat.1006146

Cited by 17 publications

(29 citation statements)

References 53 publications

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“…Both proteins are heavily phosphorylated, so it is possible that they are direct substrates of KPP1 (McAllaster et al, 2015;Urbaniak et al, 2013). In the case of TbPLK, a PEST domain that functions as a degron was recently identified in the linker domain between the N-terminal catalytic domain and the C-terminal polo-box (Hu et al, 2017). Deletion of the PEST domain in T. brucei led to the stabilization of TbPLK compared to wild type.…”

Section: Discussionmentioning

confidence: 99%

Identification of TOEFAZ1‐interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis

Hilton

Sladewski

Perry

et al. 2018

Molecular Microbiology

104

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The protist parasite Trypanosoma brucei is an obligate extracellular pathogen that retains its highly polarized morphology during cell division and has evolved a novel cytokinetic process independent of non-muscle myosin II. The polo-like kinase homolog TbPLK is essential for transmission of cell polarity during division and for cytokinesis. We previously identified a putative TbPLK substrate named Tip of the Extending FAZ 1 (TOEFAZ1) as an essential kinetoplastid-specific component of the T. brucei cytokinetic machinery. We performed a proximity-dependent biotinylation identification (BioID) screen using TOEFAZ1 as a means to identify additional proteins that are involved in cytokinesis. Using quantitative proteomic methods, we identified nearly 500 TOEFAZ1-proximal proteins and characterized 59 in further detail. Among the candidates, we identified an essential putative phosphatase that regulates the expression level and localization of both TOEFAZ1 and TbPLK, a previously uncharacterized protein that is necessary for the assembly of a new cell posterior, and a microtubule plus-end directed orphan kinesin that is required for completing cleavage furrow ingression. The identification of these proteins provides new insight into T. brucei cytokinesis and establishes TOEFAZ1 as a key component of this essential and uniquely configured process in kinetoplastids.

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Section: Discussionmentioning

confidence: 99%

Identification of TOEFAZ1‐interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis

Hilton

Sladewski

Perry

et al. 2018

Molecular Microbiology

104

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“…Transfectants were selected with blasticidin in addition to G418, hygromycin, phleomycin, and puromycin. For endogenous PTP tagging of CUL6 in cells expressing CIF1-3HA or CIF3-3HA, the pC-CUL6 -PTP-NEO vector (26) was linearized with MfeI, and transfected into the appropriate cell line.…”

Section: In Situ Epitope Tagging Of Proteinsmentioning

confidence: 99%

The trypanosome-specific protein CIF3 cooperates with the CIF1 protein to promote cytokinesis in Trypanosoma brucei

Kurasawa¹,

Hu²,

Zhou³

et al. 2018

Journal of Biological Chemistry

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Cytokinesis, the terminal step in cell division, in the protist human pathogen occurs along the longitudinal axis from the anterior tip of the new flagellum attachment zone (FAZ) toward the posterior cell tip. This process is regulated by a signaling cascade composed of the Polo-like kinase homolog TbPLK, the Aurora B kinase homolog TbAUK1, and the trypanosome-specific CIF1-CIF2 protein complex. However, the regulatory mechanism and the signaling pathway for this unusual mode of cytokinesis remain poorly understood. Here, we report another trypanosome-specific protein assembly, the CIF1-CIF3 complex, and its essential role in cytokinesis initiation. Through biochemical and genetic approaches, we demonstrate that CIF3 interacts with CIF1 in a TbPLK-dependent manner and maintains CIF1 localization at the new FAZ tip. Conversely, CIF1 maintains CIF3 stability at the new FAZ tip. We further show that TbPLK is required for CIF3 localization and that CIF3 is necessary for targeting TbAUK1 to the new FAZ tip during anaphase. These results suggest that two trypanosome-specific CIF1-containing protein complexes cooperate with the evolutionarily conserved Polo-like kinase and Aurora B kinase to promote cytokinesis in.

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“…There are a series of potential degradation motifs in the IDP, including a D-box (amino acids 366-369), a phosphodependent degron (amino acids 198-204), and a PEST domain (amino acids 399-411) (Lindon and Pines, 2004;Al-Zain et al, 2015;Li et al, 2012). Similar motifs were recently shown to control the stability of TbPLK via a ubiquitin-mediated degradation pathway (Hu et al, 2017). TOEFAZ1 lacking the IDP domain appears to persist in cells that have completed cytokinesis in a location similar to where the cleavage furrow ends, which is likely where the protein is removed and degraded (Fig.…”

Section: Discussionmentioning

confidence: 97%

A functional analysis of TOEFAZ1 uncovers protein domains essential for cytokinesis in Trypanosoma brucei

Sinclair

McAllaster

Graffenried

2017

Journal of Cell Science

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The parasite is highly polarized, including a flagellum that is attached along the cell surface by the flagellum attachment zone (FAZ). During cell division, the new FAZ positions the cleavage furrow, which ingresses from the anterior tip of the cell towards the posterior. We recently identified TOEFAZ1 (for 'Tip of the Extending FAZ protein 1') as an essential protein in trypanosome cytokinesis. Here, we analyzed the localization and function of TOEFAZ1 domains by performing overexpression and RNAi complementation experiments. TOEFAZ1 comprises three domains with separable functions: an N-terminal α-helical domain that may be involved in FAZ recruitment, a central intrinsically disordered domain that keeps the morphogenic kinase TbPLK at the new FAZ tip, and a C-terminal zinc finger domain necessary for TOEFAZ1 oligomerization. Both the N-terminal and C-terminal domains are essential for TOEFAZ1 function, but TbPLK retention at the FAZ is not necessary for cytokinesis. The feasibility of alternative cytokinetic pathways that do not employ TOEFAZ1 are also assessed. Our results show that TOEFAZ1 is a multimeric scaffold for recruiting proteins that control the timing and location of cleavage furrow ingression.

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CRL4WDR1 Controls Polo-like Kinase Protein Abundance to Promote Bilobe Duplication, Basal Body Segregation and Flagellum Attachment in Trypanosoma brucei

Cited by 17 publications

References 53 publications

Identification of TOEFAZ1‐interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis

Identification of TOEFAZ1‐interacting proteins reveals key regulators of Trypanosoma brucei cytokinesis

The trypanosome-specific protein CIF3 cooperates with the CIF1 protein to promote cytokinesis in Trypanosoma brucei

A functional analysis of TOEFAZ1 uncovers protein domains essential for cytokinesis in Trypanosoma brucei

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