hNinein is required for targeting spindle-associated protein Astrin to the centrosome during the S and G2 phases

Cheng, Tai Shan; Hsiao, Yun Ling; Lin, Ching Chih; Hsu, Che-Wei; Chang, Mau-Sun; Lee, Chu I.; Yu, Ricky Chang Tze; Huang, Chi Ying F.; Howng, Shen Long; Hong, Yi‐Ren

doi:10.1016/j.yexcr.2007.02.023

Cited by 23 publications

(27 citation statements)

References 40 publications

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“…31,32 For example, hNinein interacts with Astrin through its coiledcoil domain, which in turn is required for maintenance of centrosome/spindle pole integrity. 33 GSK3β interacts with and phosphorylates Astrin by the coiled-coil domain to target Astrin to the spindle microtubules and kinetochores. 11 Therefore, the C-terminal coiled-coil domain is able to compete effectively with endogenous Astrin protein for binding to docking proteins, and has a dominant effect on spindle organization; such a mechanism is similar to Astrin depletion.…”

Section: Discussionmentioning

confidence: 99%

Astrin regulates meiotic spindle organization, spindle pole tethering and cell cycle progression in mouse oocytes

Yuan¹,

Li²,

Liang³

et al. 2009

Cell Cycle

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Section: Discussionmentioning

confidence: 99%

Astrin regulates meiotic spindle organization, spindle pole tethering and cell cycle progression in mouse oocytes

Yuan¹,

Li²,

Liang³

et al. 2009

Cell Cycle

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“…This protein has a large coiled-coil domain at its C terminus and forms dimers or high order structures under physiological conditions (22). Interestingly, Astrin-depleted cells exhibit centrosome instability and malformation of the spindle, leading to mitosis arrest and subsequent apoptosis (20,22,23). A recent study also showed that in the absence of Astrin, kinetochore-microtubule attachments are impaired, leading to premature sister chromatid separation and centriole disengagement (24).…”

Section: Additionally Astrin Acts As a Substrate For Gsk3␤ And Is Phmentioning

confidence: 99%

“…Our recent work showed that hNinein is a GSK3␤-interacting protein, and the phosphorylation sites in hNinein are located in the C-terminal region (amino acids 1617-2090) (31,32), which overlap with the Astrin binding region (23). Therefore, Astrin may affect GSK3␤-mediated hNinein phosphorylation.…”

Section: Identification Of Astrin As a Novel Gsk3␤-interacting Protein-mentioning

confidence: 99%

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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“…Astrin, which contains two coiled coil domains in its C terminus, is associated with spindle MTs as early as prophase and functions in centrosome integrity, spindle formation prior to metaphase chromosome alignment, and chromosome segregation (28,29). Mitotic proteins, including Kinastrin/Skap (27), hNinein (30), cytoplasmic linker-associated protein-1␣ (Clasp1␣) (31), and dynein light chain 8 (32), interact with Astrin and target it to spindle poles or KTs. Several kinases also regulate the function of Astrin.…”

mentioning

confidence: 99%

Phosphorylation of Astrin Regulates Its Kinetochore Function

Chung

Park

Lee

et al. 2016

Journal of Biological Chemistry

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The error-free segregation of chromosomes, which requires the precisely timed search and capture of chromosomes by spindles during early mitotic and meiotic cell division, is responsible for genomic stability and is achieved by the spindle assembly checkpoint in the metaphase-anaphase transition. Mitotic kinases orchestrate M phase events, such as the reorganization of cell architecture and kinetochore (KT) composition with the exquisite phosphorylation of mitotic regulators, to ensure timely and temporal progression. However, the molecular mechanisms underlying the changes of KT composition for stable spindle attachment during mitosis are poorly understood. Here, we show that the sequential action of the kinase Cdk1 and the phosphatase Cdc14A control spindle attachment to KTs. During prophase, the mitotic spindle protein Spag5/Astrin is transported into centrosomes by Kinastrin and phosphorylated at Ser-135 and Ser-249 by Cdk1, which, in prometaphase, is loaded onto the spindle and targeted to KTs. We also demonstrate that Cdc14A dephosphorylates Astrin, and therefore the overexpression of Cdc14A sequesters Astrin in the centrosome and results in aberrant chromosome alignment. Mechanistically, Plk1 acts as an upstream kinase for Astrin phosphorylation by Cdk1 and targeting phospho-Astrin to KTs, leading to the recruitment of outer KT components, such as Cenp-E, and the stable attachment of spindles to KTs. These comprehensive findings reveal a regulatory circuit for protein targeting to KTs that controls the KT composition change of stable spindle attachment and chromosome integrity.The maintenance of genome integrity during mitosis is crucial for cell survival and organismal development. To generate two daughter cells with identical genetic information, each sister chromatid must be captured by spindle microtubules (MTs), 4 aligned at the mitotic equator, and segregated toward the spindle poles. Cells build a proteinaceous structure, known as the KT, at the centromere and form a bipolar spindle with an amphitelic spindle attachment to achieve accurate chromosome segregation (1). The KT is made up of protein complexes that control its localization on the chromosome, assembly, attachment to spindle MTs, and chromosome movements, such as congression and segregation (2). KTs lacking essential proteins for KT-MT attachment, such as Knl1, Mis12, and Ndc80 protein subcomplexes, are unable to attach to spindle MTs (3), which in turn results in chromosome loss and concurrent cell death (4).Entry into mitosis is controlled by the concerted action of several mitotic kinases, such as Cyclin-dependent kinase 1 (Cdk1), Polo-like kinase 1 (Plk1), and Aurora A, whose activities are regulated indirectly in a positive feedback loop (5). Plk1 activates Cdk1 by phosphorylating and activating Cdc25, which then removes inhibitory phosphorylation at the Thr-14 and Tyr-15 sites of Cdk1. Aurora A phosphorylates Thr-210 in the active loop of Plk1 with the aid of Bora (6). Cdk1 also activates Plk1 via Bora phosphorylation to promote mit...

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hNinein is required for targeting spindle-associated protein Astrin to the centrosome during the S and G2 phases

Cited by 23 publications

References 40 publications

Astrin regulates meiotic spindle organization, spindle pole tethering and cell cycle progression in mouse oocytes

Astrin regulates meiotic spindle organization, spindle pole tethering and cell cycle progression in mouse oocytes

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

Phosphorylation of Astrin Regulates Its Kinetochore Function

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