Associations between the Rho kinase-1 catalytic and PH domain regulatory unit

Craft, John W.; Zhang, Hua; Charendoff, Marc N.; Mindrebo, Jeffrey T.; Schwartz, Robert J.; Briggs, James M.

doi:10.1016/j.jmgm.2013.09.009

Cited by 5 publications

(5 citation statements)

References 28 publications

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“…The autoinhibitory region is permanently cleaved during apoptosis by caspase-3 for ROCK1 20,21 and granzyme-B for ROCK2 22 to create active ROCK kinases leading to actin-myosin contraction, membrane blebbing, and formation of apoptotic bodies. The auto-inhibition model is also supported by SPR studies showing binding between the isolated kinase and PH domains of ROCK1 23 .…”

mentioning

confidence: 71%

Ligand-induced conformational rearrangements regulate the switch between membrane-proximal and distal functions of Rho kinase 2

et al. 2020

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Rho-associated protein kinase 2 (ROCK2) is a membrane-anchored, long, flexible, multidomain, multifunctional protein. Its functions can be divided into two categories: membrane-proximal and membrane-distal. A recent study concluded that membrane-distal functions require the fully extended conformation, and this conclusion was supported by electron microscopy. The present solution small-angle X-ray scattering (SAXS) study revealed that ROCK2 population is a dynamic mixture of folded and partially extended conformers. Binding of RhoA to the coiled-coil domain shifts the equilibrium towards the partially extended state. Enzyme activity measurements suggest that the binding of natural protein substrates to the kinase domain breaks up the interaction between the N-terminal kinase and C-terminal regulatory domains, but smaller substrate analogues do not. The present study reveals the dynamic behaviour of this long, dimeric molecule in solution, and our structural model provides a mechanistic explanation for a set of membrane-proximal functions while allowing for the existence of an extended conformation in the case of membrane-distal functions.

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confidence: 71%

Ligand-induced conformational rearrangements regulate the switch between membrane-proximal and distal functions of Rho kinase 2

et al. 2020

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“…Recent electron microscopy data on full-length Rock2 have sought to challenge this view, suggesting that instead of autoinhibition Rock regulation is spatially organized within the cell at a distance governed by the length of its coiled coil region (52). This is in contrast to existing data (24,51,53) demonstrating protein-protein interactions between C-terminal regulatory sequences and the kinase domain. Furthermore, binding events within the regulatory regions of Rock have been demonstrated to alter Rock activity.…”

Section: Discussionmentioning

confidence: 91%

“…The Role of the Shrm-Rock Signaling Module in Rock Regulation-When its activity is not needed, Rock is believed to reside in an autoinhibited state in which its C-terminal PH/C1 domains directly bind to its N-terminal kinase domain (24,51). Recent electron microscopy data on full-length Rock2 have sought to challenge this view, suggesting that instead of autoinhibition Rock regulation is spatially organized within the cell at a distance governed by the length of its coiled coil region (52).…”

Section: Discussionmentioning

confidence: 99%

Structure of the Shroom-Rho Kinase Complex Reveals a Binding Interface with Monomeric Shroom That Regulates Cell Morphology and Stimulates Kinase Activity

Zalewski

Heber

et al. 2016

Journal of Biological Chemistry

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Shroom-mediated remodeling of the actomyosin cytoskeleton is a critical driver of cellular shape and tissue morphology that underlies the development of many tissues including the neural tube, eye, intestines, and vasculature. Shroom uses a conserved SD2 domain to direct the subcellular localization of Rho-associated kinase (Rock), which in turn drives changes in the cytoskeleton and cellular morphology through its ability to phosphorylate and activate non-muscle myosin II. Here, we present the structure of the human Shroom-Rock binding module, revealing an unexpected stoichiometry for Shroom in which two Shroom SD2 domains bind independent surfaces on Rock. Mutation of interfacial residues impaired Shroom-Rock binding in vitro and resulted in altered remodeling of the cytoskeleton and loss of Shroom-mediated changes in cellular morphology. Additionally, we provide the first direct evidence that Shroom can function as a Rock activator. These data provide molecular insight into the Shroom-Rock interface and demonstrate that Shroom directly participates in regulating cytoskeletal dynamics, adding to its known role in Rock localization.

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“…ROCK1/2 contains a pleckstrin homology (PH) domain that acts as a regulatory unit (37) and binds the ROCK catalytic domain thereby serving to inhibit ROCK activity. Thus, the binding of a ligand to the PH domain activates ROCK by liberating the catalytic domain (38,39). This is relevant for SHIP2 because while the ROCK PH domain can bind different phosphoinositides including PI(3,4)P2, only the SHIP2 substrate, PI(3,4,5)P3, has been shown to activate ROCK (40).…”

Section: Discussionmentioning

confidence: 99%

SHIP2 controls matrix mineralization by regulation of the RhoA/ROCK pathway and remodeling of the actin cytoskeleton

Fradet

Fitzgerald

2022

Preprint

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Mutations in INPPL1, the gene coding for SH2 Domain-Containing Inositol 5'-Phosphatase 2 (SHIP2), cause Opsismodysplasia, a severe chondrodysplasia characterized by delayed bone maturation. The mechanism by which the loss of an inositol phosphatase causes a major skeletal developmental defect is unclear. To investigate the role of SHIP2 in mineralization, the INPPL1 gene was deleted in vitro in chondrocyte and osteoblast differentiation models and the effect of the loss of SHIP2 on cell differentiation, subsequent mineralization, and on actin cytoskeleton formation was investigated. The loss of SHIP2 does not impact differentiation but, consistent with the disease phenotype, induces a significant reduction in extracellular matrix mineralization in both cell types. Absence of SHIP2 also altered the actin cytoskeleton to increase cell adhesion and focal adhesion formation. Furthermore, inhibition of actin polymerization in SHIP2-deficient cells rescued the mineralization phenotype. RhoA/ROCK, Cdc42 and Rac1 are the three main RhoGTPases responsible for actin cytoskeleton regulation in bone cells. Specific inhibitors of these RhoGTPases were used to determine the pathways involved in SHIP2-mediated mineralization. Since only the ROCK pathway inhibitor rescued the mineralization phenotype, it is concluded that SHIP2 regulates actin cytoskeleton remodeling and consequently extracellular matrix mineralization by inhibiting the RhoA/ROCK pathway.

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Associations between the Rho kinase-1 catalytic and PH domain regulatory unit

Cited by 5 publications

References 28 publications

Ligand-induced conformational rearrangements regulate the switch between membrane-proximal and distal functions of Rho kinase 2

Ligand-induced conformational rearrangements regulate the switch between membrane-proximal and distal functions of Rho kinase 2

Structure of the Shroom-Rho Kinase Complex Reveals a Binding Interface with Monomeric Shroom That Regulates Cell Morphology and Stimulates Kinase Activity

SHIP2 controls matrix mineralization by regulation of the RhoA/ROCK pathway and remodeling of the actin cytoskeleton

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