CEP70 Protein Interacts with γ-Tubulin to Localize at the Centrosome and Is Critical for Mitotic Spindle Assembly

Shi, Xingjuan; Sun, Xiaoou; Liu, Min; Li, Dengwen; Aneja, Ritu; Zhou, Jun

doi:10.1074/jbc.m111.252262

Cited by 36 publications

(43 citation statements)

References 23 publications

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“…Consistent with the previous finding [20], immunofluorescence microscopy revealed that the centrosomal localization of Cep70 was greatly dependent on its second coiled-coil domain ( Figure 4E and 4F). Importantly, the 15KR mutation significantly promoted Cep70 localization at the centrosome ( Figure 4E and 4F), suggesting that the deubiquitination of Cep70 stimulates its centrosomal localization.…”

Section: Cyld-mediated Deubiquitination Of Cep70 Promotes Its Interacsupporting

confidence: 79%

“…However, the level of Cep70 (centrosomal protein of 70 kDa), a recently identified regulator of ciliogenesis [18][19][20], at the centrosome was significantly decreased upon the depletion of CYLD, although the level of Cep70 in the total cell lysate was not affected ( Figure 3B). Immunofluorescence microscopy further confirmed that CYLD was important for Cep70 localization at the centrosome in RPE-1 cells ( Figure 3C and 3D).…”

Section: Depletion Of Cyld Impairs the Localization Of Cep70 At The Cmentioning

confidence: 89%

“…We found previously that Cep70 interacts, through its two coiled-coil domains, with γ-tubulin to localize at the centrosome [20]. It is also known that CYLD can modulate protein-protein interactions by removing polyubiquitin chains from its substrates [9].…”

Section: Cyld-mediated Deubiquitination Of Cep70 Promotes Its Interacmentioning

confidence: 99%

“…Cep70 antibody was generated as described previously [20]. Horseradish peroxidase-conjugated secondary antibodies were from Amersham Biosciences, and fluorescein-, rhodamine-, or Cy5-conjugated secondary antibodies were from Jackson ImmunoResearch Laboratories.…”

Section: Antibodies Chemicals Sirnas and Plasmidsmentioning

confidence: 99%

“…Mammalian expression plasmids for GFP-CYLD, GFP-Cep70, GST-Cep70, HA-HDAC6, and His-Myc-ubiquitin were described previously [12,20,35], and various mutants were generated by PCR and site-directed mutagenesis. MBP-Cep70 was purified from bacteria expressing the pMALp2T-Cep70 plasmid as described previously [20], with amylose resin (New England Biolabs). His-CYLD was purified from insect cells using the Bac-to-Bac baculovirus expression system (Life Technologies), with nickel resin (Bio-Rad).…”

Section: Antibodies Chemicals Sirnas and Plasmidsmentioning

confidence: 99%

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CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

et al. 2014

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Cilia are hair-like organelles extending from the cell surface with important sensory and motility functions. Ciliary defects can result in a wide range of human diseases known as ciliopathies. However, the molecular mechanisms controlling ciliogenesis remain poorly defined. Here we show that cylindromatosis (CYLD), a tumor suppressor protein harboring deubiquitinase activity, plays a critical role in the assembly of both primary and motile cilia in multiple organs. CYLD knockout mice exhibit polydactyly and various ciliary defects, such as failure in basal body anchorage and disorganization of basal bodies and axenomes. The ciliary function of CYLD is partially attributed to its deconjugation of the polyubiquitin chain from centrosomal protein of 70 kDa (Cep70), a requirement for Cep70 to interact with γ-tubulin and localize at the centrosome. In addition, CYLD-mediated inhibition of histone deacetylase 6 (HDAC6), which promotes tubulin acetylation, constitutes another mechanism for the ciliary function of CYLD. Small-molecule inhibitors of HDAC6 could partially rescue the ciliary defects in CYLD knockout mice. These findings highlight the importance of protein ubiquitination in the modulation of ciliogenesis, identify CYLD as a crucial regulator of this process, and suggest the involvement of CYLD deficiency in ciliopathies.

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Section: Cyld-mediated Deubiquitination Of Cep70 Promotes Its Interacsupporting

confidence: 79%

Section: Depletion Of Cyld Impairs the Localization Of Cep70 At The Cmentioning

confidence: 89%

Section: Cyld-mediated Deubiquitination Of Cep70 Promotes Its Interacmentioning

confidence: 99%

Section: Antibodies Chemicals Sirnas and Plasmidsmentioning

confidence: 99%

Section: Antibodies Chemicals Sirnas and Plasmidsmentioning

confidence: 99%

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CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

et al. 2014

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Activated proline‐rich tyrosine kinase 2 regulates meiotic spindle assembly in the mouse oocyte

Meng

Cui

Cheng

et al. 2017

J of Cellular Biochemistry

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Proline-rich tyrosine kinase 2 (PYK2), a member of the protein tyrosine kinase family, plays an important role in various cellular processes. PYK2 can be phosphorylated on tyrosine 402 by diverse stimuli at the cell surface, and recent studies have shown that this activated form of PYK2 is enriched in oocytes and required for fertilization. However, the subcellular localization and functions of activated PYK2 in oocytes remain elusive.In this study, we demonstrate that the localization of p-PYK2 undergoes dynamic changes during in vitro maturation of mouse oocytes. The signal of p-PYK2 is initially dispersed in the cytoplasm, but begins to decorate organized microtubules after the germinal vesicle breakdown and localizes to spindle poles at metaphase. Our data further show that p-PYK2 colocalizes with γ-tubulin from the germinal vesicle stage through the end of meiosis in mouse oocytes. Nocodazole treatment and washout experiments confirm that p-PYK2 associates with the oocyte spindle and spindle poles. Moreover, pharmacological inhibition of PYK2 activity dramatically alters the morphology of the bipolar spindle and prevents oocyte maturation. Together, these data suggest that activated PYK2 may function as a component of the microtubule organizing center to regulate spindle assembly during the meiotic process of mouse oocytes.

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Analyses stratified by maternal age and recombination further characterize genes associated with maternal nondisjunction of chromosome 21

2021

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Objective In our previous work, we performed the first genome‐wide association study to find genetic risk factors for maternal nondisjunction of chromosome 21. The objective of the current work was to perform stratified analyses of the same dataset to further elucidate potential mechanisms of genetic risk factors. Methods We focused on loci that were statistically significantly associated with maternal nondisjunction based on this same dataset in our previous study and performed stratified association analyses in seven subgroups defined by age and meiotic recombination profile. In each analysis, we contrasted a different subgroup of mothers with the same set of fathers, the mothers serving as cases (phenotype: meiotic nondisjunction of chromosome 21) and the fathers as controls. Results Our stratified analyses identified several genes whose patterns of association are consistent with generalized effects across groups, as well as other genes that are consistent with specific effects in certain groups. Conclusions While our results are epidemiological in nature and cannot conclusively prove mechanisms, we identified a number of patterns that are consistent with specific mechanisms. In many cases those mechanisms are strongly supported by available literature on the associated genes.

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CEP70 Protein Interacts with γ-Tubulin to Localize at the Centrosome and Is Critical for Mitotic Spindle Assembly

Cited by 36 publications

References 23 publications

CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

CYLD mediates ciliogenesis in multiple organs by deubiquitinating Cep70 and inactivating HDAC6

Activated proline‐rich tyrosine kinase 2 regulates meiotic spindle assembly in the mouse oocyte

Analyses stratified by maternal age and recombination further characterize genes associated with maternal nondisjunction of chromosome 21

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