Preisinger C, Short B, De Corte V, Bruyneel E, Haas A, Kopajtich R et al.. YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3zeta. J Cell Biol 164: 1009-1020

Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, Martinsried, 82152 Germany.
The Journal of Cell Biology (Impact Factor: 9.83). 04/2004; 164(7):1009-20. DOI: 10.1083/jcb.200310061
Source: PubMed


The Golgi apparatus has long been suggested to be important for directing secretion to specific sites on the plasma membrane in response to extracellular signaling events. However, the mechanisms by which signaling events are coordinated with Golgi apparatus function remain poorly understood. Here, we identify a scaffolding function for the Golgi matrix protein GM130 that sheds light on how such signaling events may be regulated. We show that the mammalian Ste20 kinases YSK1 and MST4 target to the Golgi apparatus via the Golgi matrix protein GM130. In addition, GM130 binding activates these kinases by promoting autophosphorylation of a conserved threonine within the T-loop. Interference with YSK1 function perturbs perinuclear Golgi organization, cell migration, and invasion into type I collagen. A biochemical screen identifies 14-3-3zeta as a specific substrate for YSK1 that localizes to the Golgi apparatus, and potentially links YSK1 signaling at the Golgi apparatus with protein transport events, cell adhesion, and polarity complexes important for cell migration.

Download full-text


Available from: Alexander K Haas, Jan 13, 2015
  • Source
    • "Finally, the Golgi apparatus has also been shown to play an important role in cell signaling, and its polarized position within the cell could contribute to this role. Two kinases involved in cell polarization and migration, YSK1 and MST4, were shown to localize to the Golgi apparatus through interactions with Golgi matrix protein GM130, which directly activated the kinases [95]. Ras shows diverse activation kinetics and second messenger binding partners based on its localization to the plasma membrane or Golgi apparatus, and can even be activated independently in one location or the other [96]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Cell polarization is a process of coordinated cellular rearrangements that prepare the cell for migration. GM1 is synthesized in the Golgi apparatus and localized in membrane microdomains that appear at the leading edge of polarized cells, but the mechanism by which GM1 accumulates asymmetrically is unknown. The Golgi apparatus itself becomes oriented toward the leading edge during cell polarization, which is thought to contribute to plasma membrane asymmetry. Using quantitative image analysis techniques, we measure the extent of polarization of the Golgi apparatus and GM1 in the plasma membrane simultaneously in individual cells subject to a wound assay. We find that GM1 polarization starts just 10 min after stimulation with growth factors, while Golgi apparatus polarization takes 30 min. Drugs that block Golgi polarization or function have no effect on GM1 polarization, and, conversely, inhibiting GM1 polarization does not affect Golgi apparatus polarization. Evaluation of Golgi apparatus and GM1 polarization in single cells reveals no correlation between the two events. Our results indicate that Golgi apparatus and GM1 polarization are controlled by distinct intracellular cascades involving the Ras/Raf/MEK/ERK and the PI3K/Akt/mTOR pathways, respectively. Analysis of cell migration and invasion suggest that MEK/ERK activation is crucial for two dimensional migration, while PI3K activation drives three dimensional invasion, and no cumulative effect is observed from blocking both simultaneously. The independent biochemical control of GM1 polarity by PI3K and Golgi apparatus polarity by MEK/ERK may act synergistically to regulate and reinforce directional selection in cell migration.
    Full-text · Article · Dec 2013 · PLoS ONE
  • Source
    • "Stk25 has been shown to regulate polarized migration in cultured cells, and this activity is regulated through its interactions with GM130 [20], a ubiquitously expressed Golgi-shaping protein. GM130 has been shown to regulate tethering of ER-derived vesicles with the Golgi [21,22]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Neuronal migration involves the directional migration of immature neurons. During much of the migration period these neurons are polarized with defined leading and trailing processes. Stk25 has been shown to bind to the LKB1 activator STRAD and regulate neuronal polarization and dendritogenesis in an opposing manner to Reelin-Dab1 signaling. It is not known, however, whether Stk25 controls neuronal migration, a key developmental process regulated by Reelin-Dab1 signal transduction. Here we find that while constitutive Stk25 deficiency does not lead to neuronal phenotypes, acute reduction by either Cre-mediated gene inactivation or by knockdown causes a developmental neuronal migration error. Furthermore, we find that knockdown of LKB1, STRAD and GM130, molecules that have previously been implicated with Stk25, causes similar aberrations in neuronal migration. Loss of Stk25 function early in development likely leads to functional compensation for its roles in neuronal development. Stk25 regulates neuronal positioning, possibly as part of the LKB1-STRAD-Stk25-GM130 pathway that was previously shown to be important for neuronal polarization.
    Full-text · Article · Nov 2013 · Neural Development
  • Source
    • "masks the ER localization signal in these proteins and prevents their retention in the ER ( Shikano et al . , 2005 ; Shikano et al . , 2006 ) . Cx43 is a unique membrane protein that oligomerizes in the Golgi instead of the ER ( Musil and Goodenough , 1993 ) . Indeed , a large fraction of 14 - 3 - 3h is also reported to be localized in the Golgi ( Preisinger et al . , 2004 ) . Interestingly , we observed that absence of 14 - 3 - 3h caused the accumulation of Cx43 in the Golgi , also the site where Cx43 oligomerizes to form hexameric connexon ( HC ) prior to being delivered to the plasma membrane . Moreover , FFSS promoted migration of Cx43 from the Golgi to the cell surface was accompanied by increased pr"
    [Show abstract] [Hide abstract]
    ABSTRACT: Intracellular signaling in osteocytes activated by mechanical loading is important for bone formation and remodeling. These signaling events are mediated by small modulators released from Cx43 hemichannels (HC). We have recently shown that integrin α5 senses the mechanical stimulation and induces the opening of Cx43 HC; however, the underlying mechanism is unknown. Here, we show that both Cx43 and integrin α5 interact with 14-3-3θ, and this interaction is required for the opening of Cx43 HC upon mechanical stress. Ablation of 14-3-3θ prevented the interaction between Cx43 and integrin α5, and blocked HC opening. It further decreased the transport of Cx43 and integrin α5 from the Golgi to the plasma membrane. Moreover, mechanical loading promoted the movement of Cx43 to the surface which was associated not only with an increase in 14-3-3θ levels but also its interaction with Cx43 and integrin α5. This stimulatory effect on forward transport by mechanical loading was attenuated in the absence of 14-3-3θ and majority of the Cx43 was accumulated in the Golgi. Disruption of Golgi apparatus by brefeldin A reduced the association of Cx43 and integrin α5 with 14-3-3θ, further suggesting that the interaction is likely to occur in the Golgi. Together, these results define a novel, scaffolding role of 14-3-3θ in assisting the delivery of Cx43 and integrin α5 to the plasma membrane for the formation of mechanosensitive HC in osteocytes.
    Full-text · Article · Oct 2013 · Journal of Cell Science
Show more