Characterization and Functional Aspects of Human Ninein Isoforms that Regulated by Centrosomal Targeting Signals and Evidence for Docking Sites to Direct Gamma-Tubulin

Lin, Ching‐Chih; Cheng, Tsung Chieh; Hsu, Chen-Pin; Wu, Che-Hsiang; Chang, Long‐Sen; Shen, Zhi-Shiang; Yeh, Hung-Ming; Chang, Li-Kwan; Howng, Shen-Long; Hong, Yi‐Ren

doi:10.4161/cc.5.21.3404

Cited by 25 publications

(24 citation statements)

References 35 publications

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“…5 and 6). This is consistent with several studies showing a role for GSK3␤ in chromosomal alignment and microtubule organization at the centrosome (18,19,28,40,41). It is, therefore, reasonable that GSK3␤ kinase activity is indeed as an important factor in mitotic spindle regulation.…”

Section: Gsk3␤-astrin Complex Exclusively Localizes On Spindle Microtsupporting

confidence: 79%

Glycogen Synthase Kinase 3β Interacts with and Phosphorylates the Spindle-associated Protein Astrin

Cheng

Hsiao

Lin

et al. 2008

Journal of Biological Chemistry

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Emerging evidence shows that glycogen synthase kinase 3␤ (GSK3␤) is involved in mitotic division and that inhibiting of GSK3␤ kinase activity causes defects in spindle microtubule length and chromosome alignment. However, the purpose of GSK3␤ involvement in spindle microtubule assembly and accurate chromosome segregation remains obscure. Here, we report that GSK3␤ interacts with the spindle-associated protein Astrin both in vitro and in vivo. Additionally, Astrin acts as a substrate for GSK3␤ and is phosphorylated at Thr-111, Thr-937 ((S/T)P motif) and Ser-974/Thr-978 ((S/T)XXX(S/T)-p motif; p is a phosphorylatable residue). Inhibition of GSK3␤ impairs spindle and kinetochore accumulation of Astrin and spindle formation at mitosis, suggesting that Astrin association with the spindle microtubule and kinetochore may be dependent on phosphorylation by GSK3␤. Conversely, depletion of Astrin by small interfering RNA has no detectable influence on the localization of GSK3␤. Interestingly, in vitro assays demonstrated that Astrin enhances GSK3␤-mediated phosphorylation of other substrates. Moreover, we showed that coexpression of Astrin and GSK3␤ differentially increases GSK3␤-mediated Tau phosphorylation on an unprimed site. Collectively, these data indicate that GSK3␤ interacts with and phosphorylates the spindle-associated protein Astrin, resulting in targeting Astrin to the spindle microtubules and kinetochores. In turn, the GSK3␤-Astrin complex may also facilitate further physiological and pathological phosphorylation.Glycogen synthase kinase 3 (GSK3), 2 a serine/threonine kinase active in several signaling pathways, is involved in the regulation of cell fate, including Wnt and Hedgehog signal transduction, protein synthesis, glycogen metabolism, mitosis and apoptosis (1-4). GSK3 has two structurally similar isoforms in mammals, GSK3␣ and GSK3␤, that are ubiquitously expressed and differ in their N-and C-terminal regions (2, 5). Earlier reports indicated that not only are the developmental profiles of GSK3␣ and GSK3␤ expression different but also the regulation and functions of these two proteins are not always identical (6 -9). Factors known to influence the functions of GSK3␤ include the phosphorylation of GSK3␤ itself, the subcellular localization of GSK3␤, the protein-protein interaction of GSK3␤, and the phosphorylation state of GSK3␤ substrates (1, 3, 4, 10). Insulin-mediated inhibition of GSK3 was mediated through a phosphorylation-dependent mechanism, with the phosphorylation at position Ser-21 and Ser-9 in GSK3␣ and GSK3␤, respectively (11).GSK3␤ also has a preference for pre-phosphorylated substrates, recognizing the consensus sequence (S/T)XXX(S/T)-p. In this sequence the first S/T residue is the target for GSK3␤ phosphorylation, X is any amino acid, P denotes the phosphorylatable residue, and the S/T located at the C terminus is the phosphorylation priming site (12). On the other hand, a number of proteins, including Axin and Tau, are phosphorylated by GSK3␤ without pre-phosphorylation. Recombinant Tau...

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Section: Gsk3␤-astrin Complex Exclusively Localizes On Spindle Microtsupporting

confidence: 79%

Glycogen Synthase Kinase 3β Interacts with and Phosphorylates the Spindle-associated Protein Astrin

Cheng

Hsiao

Lin

et al. 2008

Journal of Biological Chemistry

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“…Depending on the level of expression, GFP-C-ninein also accumulated in small dots in the cytoplasm, as described previously (30). At high levels of expression, GFP-C-ninein formed aggregates in the cytoplasm, as described previously (56). To test whether overexpression of GFPninein WT and GFP-C-ninein alters the MT cytoskeleton, the cells showing a modest level of expression were immunolabeled with ␤-tubulin.…”

Section: Jak2 Tyrosyl Phosphorylates the N Terminus Of Ninein (Amino mentioning

confidence: 99%

JAK2 Tyrosine Kinase Phosphorylates and Is Negatively Regulated by Centrosomal Protein Ninein

Jay

Hammer

Nestor-Kalinoski

et al. 2015

Molecular and Cellular Biology

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JAK2 is a cytoplasmic tyrosine kinase critical for cytokine signaling. In this study, we have identified a novel centrosome-associated complex containing ninein and JAK2. We have found that active JAK2 localizes around the mother centrioles, where it partly colocalizes with ninein, a protein involved in microtubule (MT) nucleation and anchoring. We demonstrated that JAK2 is an important regulator of centrosome function. Depletion of JAK2 or use of JAK2-null cells causes defects in MT anchoring and increased numbers of cells with mitotic defects; however, MT nucleation is unaffected. We showed that JAK2 directly phosphorylates the N terminus of ninein while the C terminus of ninein inhibits JAK2 kinase activity in vitro. Overexpressed wild-type (WT) or C-terminal (amino acids 1179 to 1931) ninein inhibits JAK2. This ninein-dependent inhibition of JAK2 significantly decreases prolactin-and interferon gamma (IFN-␥)-induced tyrosyl phosphorylation of STAT1 and STAT5. Downregulation of ninein enhances JAK2 activation. These results indicate that JAK2 is a novel member of centrosome-associated complex and that this localization regulates both centrosomal function and JAK2 kinase activity, thus controlling cytokine-activated molecular pathways. JAK2 is a tyrosine kinase that is activated by two-thirds of the cytokine/hematopoietin receptor superfamily. JAK2 activation initiates a variety of downstream signaling events that lead to diverse physiological responses to cytokines, including regulation of body growth, hematopoiesis, satiety, lactation, and various immune functions. Upon activation, receptor-bound JAK2 phosphorylates specific tyrosine residues of its downstream targets, activating cell survival/proliferation-promoting signaling pathways (1). One target of JAK2 is a family of transcription factors termed signal transducers and activators of transcription (STATs). STATs exist within the cytoplasm in a latent or inactive state; they are recruited by cytokine receptor complexes through interaction between the receptor or JAK phosphotyrosines and the Src homology 2 (SH2) domain of the STAT protein. Several additional kinase cascades may be activated by JAK2, including phosphoinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways. Three mechanisms of JAK2 activation at the genetic level have been implicated in hematologic malignancies: point mutations, translocations, and amplifications (reviewed in reference 2). Mutations within the pseudokinase domain of JAK2 were detected in a spectrum of myeloproliferative diseases. A V617F gain-of-function point mutation is the cause of ϳ90% of all polycythemia vera cases and ϳ50% of all essential thrombocythemia and primitive myelofibrosis cases (3, 4). Several additional mutations leading to constitutively active JAK2 have been identified recently in patients, including a JAK2 K539L mutant. A T875N substitution in JAK2 was also identified in megakaryoblastic myeloid leukemia cells. In a patient with trisomy 21 and B cell precursor acute lymphoblastic l...

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“…1A) (Joberty et al, 2000). It also does not contain any known centrosomal targeting motifs, such as the PACT domain, which is present in AKAP450 and pericentrin (Gillingham and Munro, 2000), or sequence homology with other mother centriolespecific protein, such as ninein or Cep170 (Guarguaglini et al, 2005;Lin et al, 2006;Hüber et al, 2008). Thus, the Par6c Cterminus appears to contain a novel mother centriole-specific sorting signal.…”

Section: Discussionmentioning

confidence: 99%

“…This older centriole is decorated with functionally important protein appendages (Bornens, 2002;Bornens and Piel, 2002). Subdistal appendages recruit proteins, such as Cep170 and ninein, which control the anchoring of interphase microtubules (Guarguaglini et al, 2005;Lin et al, 2006). Distal appendages, in contrast, contain Cep164, which is proposed to play a role in the docking of the mother centriole with the plasma membrane and is required for ciliogenesis (Graser et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Par6γ is at the mother centriole and controls centrosomal protein composition through a Par6α-dependent pathway

Dormoy

Tormanen

Sütterlin

2012

Journal of Cell Science

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SummaryThe centrosome contains two centrioles that differ in age, protein composition and function. This non-membrane bound organelle is known to regulate microtubule organization in dividing cells and ciliogenesis in quiescent cells. These specific roles depend on protein appendages at the older, or mother, centriole. In this study, we identified the polarity protein partitioning defective 6 homolog gamma (Par6c) as a novel component of the mother centriole. This specific localization required the Par6c C-terminus, but was independent of intact microtubules, the dynein/dynactin complex and the components of the PAR polarity complex. Par6c depletion resulted in altered centrosomal protein composition, with the loss of a large number of proteins, including Par6a and p150Glued , from the centrosome. As a consequence, there were defects in ciliogenesis, microtubule organization and centrosome reorientation during migration. Par6c interacted with Par3 and aPKC, but these proteins were not required for the regulation of centrosomal protein composition. Par6c also associated with Par6a, which controls protein recruitment to the centrosome through p150 Glued . Our study is the first to identify Par6c as a component of the mother centriole and to report a role of a mother centriole protein in the regulation of centrosomal protein composition.

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Characterization and Functional Aspects of Human Ninein Isoforms that Regulated by Centrosomal Targeting Signals and Evidence for Docking Sites to Direct Gamma-Tubulin

Cited by 25 publications

References 35 publications

Glycogen Synthase Kinase 3β Interacts with and Phosphorylates the Spindle-associated Protein Astrin

Glycogen Synthase Kinase 3β Interacts with and Phosphorylates the Spindle-associated Protein Astrin

JAK2 Tyrosine Kinase Phosphorylates and Is Negatively Regulated by Centrosomal Protein Ninein

Par6γ is at the mother centriole and controls centrosomal protein composition through a Par6α-dependent pathway

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