FAM83D directs protein kinase CK1α to the mitotic spindle for proper spindle positioning

Fulcher, Luke J.; He, Zhengcheng; Mei, Lin; Macartney, Thomas; Wood, Nicola T.; Prescott, Alan R.; Whigham, Arlene; Varghese, Joby; Gourlay, Robert; Ball, Graeme; Clarke, Rosie G; Campbell, David G.; Maxwell, Christopher A.; Sapkota, Gopal P.

doi:10.1101/480616

Cited by 11 publications

(31 citation statements)

References 47 publications

(65 reference statements)

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“…This is consistent with the nucleolus as a site of mitotic spindle elongation and thus of chromosome segregation [41] [64]. Our data suggest that the nucleolar pool of CK1.2 might be redistributed onto the mitotic spindle during mitosis, and by analogy to human CK1α might be involved in spindle positioning [65]. The redistribution of nucleolar proteins has been described for other kinetoplastid proteins such as TbNOP86, a protein potentially involved in chromosome segregation in T. brucei [43] and LdTWF, an actin-binding protein that controls mitotic spindle elongation in Leishmania [41].…”

Section: Discussionsupporting

confidence: 84%

The low complexity regions in the C-terminus are essential for the subcellular localisation of Leishmania casein kinase 1 but not for its activity

Martel

Pine

Bartsch

et al. 2020

Preprint

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Casein Kinase 1 (CK1) family members are serine/threonine protein kinases ubiquitously expressed in eukaryotic organisms. They are involved in a wide range of important cellular processes, such as membrane trafficking, or vesicular transport in organisms from yeast to humans. Due to its broad spectrum of action, CK1 activity and expression is tightly regulated by a number of mechanisms, including subcellular sequestration. Defects in CK1 regulation, localisation or the introduction of mutations in the CK1 coding sequence are often associated with important diseases such as cancer. Increasing evidence suggest that the manipulation of host cell CK1 signalling pathways by intracellular pathogens, either by exploiting the host CK1 or by exporting the CK1 of the pathogen into the host cell may play an important role in infectious diseases. Leishmania CK1.2 is essential for parasite survival and released into the host cell, playing an important role in host pathogen interactions. Although Leishmania CK1.2 has dual role in the parasite and in the host cell, nothing is known about its parasitic localisation and organelle-specific functions. In this study, we show that CK1.2 is a ubiquitous kinase, which is present in the cytoplasm, associated to the cytoskeleton and localised to various organelles, indicating potential roles in kinetoplast and nuclear segregation, as well as ribosomal processing and motility. Furthermore, using truncated mutants, we show for the first time that the two low complexity regions (LCR) present in the C-terminus of CK1.2 are essential for the subcellular localisation of CK1.2 but not for its kinase activity, whereas the deletion of the N-terminus leads to a dramatic decrease in CK1.2 abundance. In conclusion, our data on the localisation and regulation of Leishmania CK1.2 contribute to increase the knowledge on this essential kinase and get insights into its role in the parasite.

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Section: Discussionsupporting

confidence: 84%

The low complexity regions in the C-terminus are essential for the subcellular localisation of Leishmania casein kinase 1 but not for its activity

Martel

Pine

Bartsch

et al. 2020

Preprint

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“…Retroviruses were produced using the following constructs; pBABED.puro FAM83F WT (DU37979), pBABED.puro FAM83F F284A/F288A (DU28196), pBABED.puro FLAG CRBN (DU54685) and pBABED.puro FLAG CRBN V388I (DU64137). Retroviruses were produced by transfecting HEK293-FT cells as previously described (16). Briefly, 6 g pBabe plasmid, 3.8 g pCMV5-GAG/Pol (Clontech), 2.2 g pCMV5-VSV-G (Clontech) were diluted in 600 l OptiMem (Gibco) and 24 l PEI (1 mg/ml) was added.…”

Section: Retroviral Transductionsmentioning

confidence: 99%

“…All FAM83 proteins interact with CK1, while FAM83A, B, E and H also interact with CK1 and  (14). Whilst the FAM83 family remains largely uncharacterised, roles for specific FAM83-CK1 complexes have been established in mitosis (16,17) and canonical Wnt signalling (18)(19)(20). Given the reports of IMiD-induced degradation of CK1, we sought to establish the effect of IMiDs on the stability of FAM83 proteins and different FAM83-CK1 complexes.…”

Section: Introductionmentioning

confidence: 99%

IMiDs induce FAM83F degradation via an interaction with CK1α to attenuate Wnt signalling

Sapkota

Dunbar

Macartney

2020

Preprint

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Immunomodulatory imide drugs (IMiDs) bind CRBN, a substrate receptor of the Cul4A E3 ligase complex, enabling neo-substrate recruitment and degradation via the ubiquitin-proteasome system. Here, we report FAM83F as such a neo-substrate. We recently showed that the eight FAM83 proteins (A-H) interact with members of the serine/threonine protein kinase CK1 family, to regulate their subcellular distribution and distinct biological roles. CK1α is a well-established IMiD neo-substrate and we demonstrate here that IMiD-induced FAM83F degradation requires its association with CK1α. Despite all FAM83 proteins interacting with CK1α, no other FAM83 protein is degraded by IMiDs. FAM83F is localised to the plasma membrane, and consistent with this, IMiD treatment results in depletion of both FAM83F and CK1α levels from the plasma membrane. We have recently identified FAM83F as a mediator of the canonical Wnt signalling pathway. The IMiD-induced degradation of FAM83F attenuated Wnt signalling in colorectal cancer cells and removed CK1α from the plasma membrane, mirroring the phenotypes observed with genetic ablation of FAM83F. Intriguingly, in many cancer cell lines, IMiD-induced degradation of CK1α is only modest and incomplete. In line with this observation, the expression of FAM83G, which also binds to CK1α, appears to attenuate the IMiD-induced degradation of CK1α, suggesting a protective role for FAM83G on CK1α. Our findings reveal that the efficiency of target protein degradation by IMiDs, and perhaps other degraders such as PROTACs, relies on the nature of the inherent multiprotein complex in which the target protein exists. Our findings unearth opportunities for developing degraders to target specific protein complexes.

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“…Interestingly, the expression of these two genes was decreased in HCV-cirrhosis tissues as compared to normal tissues, and was gradually increased as HCV-HCC developed. CDKN3 and FAM83D are the important regulators of cell cycle that however have received little attention, especially in HCV-HCC occurrence and progression on the background of HCV-cirrhosis [24][25][26]. We hypothesized that CDKN3 and FAM83D played a more signi cant role in HCV-HCC occurrence and progression and might be better biomarkers for early detection of HCV-HCC on the background of HCV-cirrhosis.…”

Section: Resultsmentioning

confidence: 99%

Identification of Key Genes in HCV-induced HCC at Early Stage on the Background of HCV-cirrhosis

nie

et al. 2020

Preprint

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Background: Hepatocellular carcinoma (HCC) is one of the most common and deadly malignant tumors worldwide. Hepatitis C virus (HCV)-induced cirrhosis (HCV-cirrhosis) is one of the leading causes of HCC occurrence. Due to the lack of effective biomarkers, most patients with HCV-induced HCC (HCV-HCC) are at advanced stages when they are first diagnosed. Our study aims to employ transcriptomic profiling dataset to identify potential biomarkers that can predict HCV-HCC at early stage on the background of HCV-cirrhosis. Methods: The dataset incorporating HCV-cirrhosis and HCV-HCC subjects at different stages from Gene Expression Omnibus was analyzed to identify gene signature-related to HCV-HCC on the background of HCV-cirrhosis. Multiple-genes based risk score-related prediction model was established to predict HCV-HCC samples at different stages, and receiver-operating characteristic (ROC) curve analysis was used to select the best cutoff value of risk scores for prediction and to evaluate the prediction model. Samples with risk scores higher than cutoff value were defined as high score group.Results: Highly clustered 20 genes were identified and were all gradually increased as HCV-HCC progressed from HCV-cirrhosis to very advanced HCC. Compared with HCV-cirrhosis, the prevalence of HCV-HCC at any of the stages in high score group was 100%. The risk score-related prediction model based on the 20 genes-biomarker got the accuracy of over 95.2 % with area under ROC (AUC) over 0.94. To implement the biomarker easily in real life and make it economical, we tried to limit the gene numbers. Interestingly, CDKN3 and FAM83D were significantly decreased in HCV-cirrhosis tissues as compared to normal tissues and then increased as HCV-HCC progressed. The results were attractive that the prediction model based on this 2 genes-biomarker indicated that prevalence of HCV-HCC in high score group was still 100% and got the predictive accuracy of over 95.2 % with AUC over 0.96, revealing even a better performance.Conclusion: Our study indicated a 2-genes-based biomarker that could identify HCV-HCC at earlier stages on the background of HCV-cirrhosis, which might be a promising biomarker for early diagnosis of HCV-HCC and potential novel treatment target.

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FAM83D directs protein kinase CK1α to the mitotic spindle for proper spindle positioning

Cited by 11 publications

References 47 publications

The low complexity regions in the C-terminus are essential for the subcellular localisation of Leishmania casein kinase 1 but not for its activity

The low complexity regions in the C-terminus are essential for the subcellular localisation of Leishmania casein kinase 1 but not for its activity

IMiDs induce FAM83F degradation via an interaction with CK1α to attenuate Wnt signalling

Identification of Key Genes in HCV-induced HCC at Early Stage on the Background of HCV-cirrhosis

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