RhoGDI facilitates geranylgeranyltransferase-I-mediated RhoA prenylation

Tnimov, Zakir; Abankwa, Daniel; Alexandrov, Kirill

doi:10.1016/j.bbrc.2014.09.024

Cited by 7 publications

(13 citation statements)

References 25 publications

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“…GGTase-I has been demonstrated to prenylate RhoA in cells and in vitro. 20,21 As a GTPase, RhoA cycles between three states: nucleotide free, GTP-, or GDP-bound; however, the effect of RhoA nucleotide status on GGTase-I reactivity has not yet been analyzed. Therefore, we measured the influence of RhoA nucleotide status on prenylation rates by assaying the incorporation of radiolabeled geranylgeranyl catalyzed by GGTase-I.…”

Section: Resultsmentioning

confidence: 99%

“…RhoA is a small GTPase that contains a C-terminal CaaL motif and an upstream polybasic region (PBR, Figure A). GGTase-I has been demonstrated to prenylate RhoA in cells and in vitro . , As a GTPase, RhoA cycles between three states: nucleotide-free, GTP-bound, or GDP-bound. However, the effect of RhoA nucleotide status on GGTase-I reactivity has not yet been analyzed.…”

Section: Resultsmentioning

confidence: 99%

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SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent

et al. 2018

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Protein prenylation involves the attachment of a hydrophobic isoprenoid moiety to the C-terminus of proteins. Several small GTPases, including members of the Ras and Rho subfamilies, require prenylation for their normal and pathological functions. Recent work has suggested that SmgGDS proteins regulate the prenylation of small GTPases in vivo. Using RhoA as a representative small GTPase, we directly test this hypothesis using biochemical assays and present a mechanism describing the mode of prenylation regulation. SmgGDS-607 completely inhibits RhoA prenylation catalyzed by protein geranylgeranyltransferase I (GGTase-I) in an in vitro radiolabel incorporation assay. SmgGDS-607 inhibits prenylation by binding to and blocking access to the C-terminal tail of the small GTPase (substrate sequestration mechanism) rather than via inhibition of the prenyltransferase activity. The reactivity of GGTase-I with RhoA is unaffected by addition of nucleotides. In contrast, the affinity of SmgGDS-607 for RhoA varies with the nucleotide bound to RhoA; SmgGDS-607 has a higher affinity for RhoA-GDP compared to RhoA-GTP. Consequently, the prenylation blocking function of SmgGDS-607 is regulated by the bound nucleotide. This work provides mechanistic insight into a novel pathway for the regulation of small GTPase protein prenylation by SmgGDS-607 and demonstrates that peptides are a good mimic for full-length proteins when measuring GGTase-I activity.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent

et al. 2018

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“…It has been demonstrated that the isoprenylation process in cells can be regulated by GDIs [ 69 ]. GDI mediates the release of RHO GTPases from the membrane, maintains them in an inactivated state, and safeguards them against degradation or nonspecific activation by RHOGEFs [ 25 , 29 , 30 ].…”

Section: Regulation Of Rho Family Gtpasesmentioning

confidence: 99%

The RHO Family GTPases: Mechanisms of Regulation and Signaling

Mosaddeghzadeh

Ahmadian

2021

Cells

177

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Much progress has been made toward deciphering RHO GTPase functions, and many studies have convincingly demonstrated that altered signal transduction through RHO GTPases is a recurring theme in the progression of human malignancies. It seems that 20 canonical RHO GTPases are likely regulated by three GDIs, 85 GEFs, and 66 GAPs, and eventually interact with >70 downstream effectors. A recurring theme is the challenge in understanding the molecular determinants of the specificity of these four classes of interacting proteins that, irrespective of their functions, bind to common sites on the surface of RHO GTPases. Identified and structurally verified hotspots as functional determinants specific to RHO GTPase regulation by GDIs, GEFs, and GAPs as well as signaling through effectors are presented, and challenges and future perspectives are discussed.

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“…The shuttling process, which considerably differs from the KRAS4B Far -PDEδ [28][29][30][31], involves the extraction of RHO GTPases from donor membranes, formation of cytosolic GDI-RHO GTPase complexes and delivery of RHO GTPases to the target membranes [13,27]. Accordingly, it has been proposed that GDI regulates the isoprenylation process in the cell [32]. GDI is known to extract RHO GTPases from the membrane, maintain them in an inactivated state, and protect them from both degradation and unspecific activation by RHO-specific GEFs [13,33,34].…”

Section: Introductionmentioning

confidence: 99%

Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions

Mosaddeghzadeh

Jasemi

Majolée

et al. 2021

IJMS

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Three decades of research have documented the spatiotemporal dynamics of RHO family GTPase membrane extraction regulated by guanine nucleotide dissociation inhibitors (GDIs), but the interplay of the kinetic mechanism and structural specificity of these interactions is as yet unresolved. To address this, we reconstituted the GDI-controlled spatial segregation of geranylgeranylated RHO protein RAC1 in vitro. Various biochemical and biophysical measurements provided unprecedented mechanistic details for GDI function with respect to RHO protein dynamics. We determined that membrane extraction of RHO GTPases by GDI occurs via a 3-step mechanism: (1) GDI non-specifically associates with the switch regions of the RHO GTPases; (2) an electrostatic switch determines the interaction specificity between the C-terminal polybasic region of RHO GTPases and two distinct negatively-charged clusters of GDI1; (3) a non-specific displacement of geranylgeranyl moiety from the membrane sequesters it into a hydrophobic cleft, effectively shielding it from the aqueous milieu. This study substantially extends the model for the mechanism of GDI-regulated RHO GTPase extraction from the membrane, and could have implications for clinical studies and drug development.

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RhoGDI facilitates geranylgeranyltransferase-I-mediated RhoA prenylation

Cited by 7 publications

References 25 publications

SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent

SmgGDS-607 Regulation of RhoA GTPase Prenylation Is Nucleotide-Dependent

The RHO Family GTPases: Mechanisms of Regulation and Signaling

Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions

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