Interaction characteristics of Plexin-B1 with Rho family proteins

Fansa, Eyad K.; Dvorský, Radovan; Zhang, Si-Cai; Fiegen, Dennis; Ahmadian, Mohammad Réza

doi:10.1016/j.bbrc.2013.04.012

Cited by 16 publications

(20 citation statements)

References 32 publications

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“…A series of later studies, through genetic association, yeast two-hybrid and biochemical co-purification, found that the insertion region between the two GAP-homology segments binds directly to some Rho family GTPases in a GTP-dependent manner (Driessens et al, 2001; Hu et al, 2001; Oinuma et al, 2003; Rohm et al, 2000; Turner et al, 2004; Vikis et al, 2000; Zanata et al, 2002). A systematic survey has shown that the RBD of PlexinB1 binds Rac1, Rac2, Rac3, Rnd1, Rnd2, Rnd3 and RhoD, but not RhoA, Cdc42, RhoG or Rif (Fansa et al, 2013). …”

Section: Regulation Of Plexin By Rhogtpasesmentioning

confidence: 99%

“…However, the plexin/RhoGTPase interactions are rather weak, with the dissociation constant in the 3-40 µM range (Fansa et al, 2013; Hota and Buck, 2009; Tong et al, 2007; Tong et al, 2009; Wang et al, 2011). It seems unlikely such a weak interaction can effectively compete with the RhoGTPase/effector interactions, which are often tighter.…”

Section: Regulation Of Plexin By Rhogtpasesmentioning

confidence: 99%

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Structural mechanisms of plexin signaling

Pascoe

Wang

Zhang

2015

Progress in Biophysics and Molecular Biology

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Signaling through plexin, the major cell surface receptor for semaphorin, plays critical roles in regulating processes such as neuronal axon guidance, angiogenesis and immune response. Plexin is normally kept inactive in the absence of semaphorin. Upon binding of semaphorin to the extracellular region, plexin is activated and transduces signal to the inside of the cell through its cytoplasmic region. The GTPase Activating Protein (GAP) domain in the plexin cytoplasmic region mediates the major intracellular signaling pathway. The substrate specificity and regulation mechanisms of the GAP domain have only been revealed recently. Many intracellular proteins serve as either upstream regulators or downstream transducers by directly interacting with plexin. The mechanisms of action for some of these proteins also start to emerge from recent studies. We review here these advances in the mechanistic understanding of plexin intracellular signaling from a structural perspective.

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Section: Regulation Of Plexin By Rhogtpasesmentioning

confidence: 99%

Section: Regulation Of Plexin By Rhogtpasesmentioning

confidence: 99%

Structural mechanisms of plexin signaling

Pascoe

Wang

Zhang

2015

Progress in Biophysics and Molecular Biology

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Section: Introductionmentioning

confidence: 99%

“…1A) (Fansa et al, 2013; Pascoe et al, 2015). Several Rho GTPases have been reported to bind to the RBD of plexin-B1 in addition to Rnd1, including Rac1, RhoD, Rnd2 and Rnd3 (Fansa et al, 2013). Rnd proteins can also bind to other plexins, such as plexin-A1 and plexin-D1 (Rohm et al, 2000; Uesugi et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Rnd3-induced cell rounding requires interaction with Plexin-B2

McColl

Garg

Riou

et al. 2016

Journal of Cell Science

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Rnd proteins are atypical members of the Rho GTPase family that induce actin cytoskeletal reorganization and cell rounding. Rnd proteins have been reported to bind to the intracellular domain of several plexin receptors, but whether plexins contribute to the Rnd-induced rounding response is not known. Here we show that Rnd3 interacts preferentially with plexin-B2 of the three plexin-B proteins, whereas Rnd2 interacts with all three B-type plexins, and Rnd1 shows only very weak interaction with plexin-B proteins in immunoprecipitations. Plexin-B1 has been reported to act as a GAP for R-Ras and/or Rap1 proteins. We show that all three plexin-B proteins interact with R-Ras and Rap1, but Rnd proteins do not alter this interaction or R-Ras or Rap1 activity. We demonstrate that plexin-B2 promotes Rnd3-induced cell rounding and loss of stress fibres, and enhances the inhibition of HeLa cell invasion by Rnd3. We identify the amino acids in Rnd3 that are required for plexin-B2 interaction, and show that mutation of these amino acids prevents Rnd3-induced morphological changes. These results indicate that plexin-B2 is a downstream target for Rnd3, which contributes to its cellular function.

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“…The protein solution was concentrated and further purified on a Superdex 75 column (10/300 GL, GE-Healthcare, Uppsala, Sweden) with 50 mM HEPES-NaOH pH 7.4, 150 mM NaCl, 3 mM DTT, 5 mM MgCl 2 and 0.5% (w/v) Na-cholate as buffer system. GppNHp-bound Rac1 proteins as well as human GDI1, RacGEFs, Pak1, full length and C-terminal truncated Rac1 proteins were prepared from E. coli as described previously [11], [43].…”

Section: Methodsmentioning

confidence: 99%

Liposome Reconstitution and Modulation of Recombinant Prenylated Human Rac1 by GEFs, GDI1 and Pak1

et al. 2014

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Small Rho GTPases are well known to regulate a variety of cellular processes by acting as molecular switches. The regulatory function of Rho GTPases is critically dependent on their posttranslational modification at the carboxyl terminus by isoprenylation and association with proper cellular membranes. Despite numerous studies, the mechanisms of recycling and functional integration of Rho GTPases at the biological membranes are largely unclear. In this study, prenylated human Rac1, a prominent member of the Rho family, was purified in large amount from baculovirus-infected Spodoptera frugiperda insect cells using a systematic detergent screening. In contrast to non-prenylated human Rac1 purified from Escherichia coli, prenylated Rac1 from insect cells was able to associate with synthetic liposomes and to bind Rho-specific guanine nucleotide dissociation inhibitor 1 (GDI1). Subsequent liposome reconstitution experiments revealed that GDI1 efficiently extracts Rac1 from liposomes preferentially in the inactive GDP-bound state. The extraction was prevented when Rac1 was activated to its GTP-bound state by Rac-specific guanine nucleotide exchange factors (GEFs), such as Vav2, Dbl, Tiam1, P-Rex1 and TrioN, and bound by the downstream effector Pak1. We found that dissociation of Rac1-GDP from its complex with GDI1 strongly correlated with two distinct activities of especially Dbl and Tiam1, including liposome association and the GDP/GTP exchange. Taken together, our results provided first detailed insights into the advantages of the in vitro liposome-based reconstitution system to study both the integration of the signal transducing protein complexes and the mechanisms of regulation and signaling of small GTPases at biological membranes.

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Interaction characteristics of Plexin-B1 with Rho family proteins

Cited by 16 publications

References 32 publications

Structural mechanisms of plexin signaling

Structural mechanisms of plexin signaling

Rnd3-induced cell rounding requires interaction with Plexin-B2

Liposome Reconstitution and Modulation of Recombinant Prenylated Human Rac1 by GEFs, GDI1 and Pak1

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