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p63RhoGEF and GEFT are Rho-specific guanine nucleotide exchange factors encoded by the same gene - Naunyn-Schmiedeberg's Archives of Pharmacology

  • ️Wieland, Thomas
  • ️Wed Apr 07 2004

Abstract

Activation of Rho GTPases, which play pivotal roles in diverse cellular functions, is catalysed by specific guanine nucleotide exchange factors (GEFs). We and others (Souchet et al. (2002)) independently cloned a human cDNA encoding a 580 aa protein (p63RhoGEF), which contains a tandem of Dbl homology and pleckstrin homology domains typical for RhoGEFs. In accordance with Souchet et al., recombinant p63RhoGEF interacted with and catalysed GDP/GTP exchange at RhoA, but not Rac1 or Cdc42. Recently, an N-terminally truncated form of p63RhoGEF, termed GEFT, was described as a Rac/Cdc42-specific GEF (Guo et al. 2003). As judged by RT-PCR with specific primers, we were able to detect mRNA variants encoding p63RhoGEF and GEFT within several tissues and cell lines. Apparently, they co-exist within one cell and are derived from the same gene. When expressed in human embryonic kidney cells, both p63RhoGEF and GEFT caused activation of RhoA, but not Rac1 or Cdc42, and induced serum response factor-mediated gene transcription, which was fully blunted by the Rho-inactivating C3 transferase. In line with these data, expression of either p63RhoGEF or GEFT in J82 human bladder carcinoma cells induced the formation of actin stress fibres. We therefore conclude that p63RhoGEF and GEFT are apparently isoforms derived from the same gene and that GEFT, similar to p63RhoGEF, activates RhoA in several cell types.

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References

  • Aghazadeh B, Lowry WE, Huang XY, Rosen MK (2000) Structural basis for relief of autoinhibition of the Dbl homology domain of proto-oncogene Vav by tyrosine phosphorylation. Cell 102:625–633

    CAS  PubMed  Google Scholar 

  • Aktories K, Schmidt G, Just I (2000) Rho GTPases as targets of bacterial protein toxins. Biol Chem 381:421–426

    CAS  PubMed  Google Scholar 

  • Benard V, Bokoch GM (2002) Assay of Cdc42, Rac, and Rho GTPase activation by affinity methods. Methods Enzymol 345:349–359

    PubMed  Google Scholar 

  • Blomquist A, Schwörer G, Schablowski H, Psoma A, Lehnen M, Jakobs KH, Rümenapp U (2000) Identification and characterization of a novel Rho-specific guanine nucleotide exchange factor. Biochem J 352:319–325

    Article  CAS  PubMed  Google Scholar 

  • Debant A, Serra-Pages C, Seipel K, O’Brien S, Tang M, Park SH, Streuli M (1996) The multidomain protein Trio binds the LAR transmembrane tyrosine phosphatase, contains a protein kinase domain, and has separate Rac-specific and Rho-specific guanine nucleotide exchange factor domains. Proc Natl Acad Sci USA 93:5466–5471

    Article  CAS  PubMed  Google Scholar 

  • Guo X, Stafford LJ, Bryan B, Xia C, Ma W, Wu X, Liu D, Songyang Z, Liu M (2003) A Rac/Cdc42-specific exchange factor, GEFT, induces cell proliferation, transformation, and migration. J Biol Chem 278:13207–13215

    Article  CAS  PubMed  Google Scholar 

  • Hart MJ, Eva A, Zangrilli D, Aaronson SA, Evans T, Cerione RA, Zheng Y (1994) Cellular transformation and guanine nucleotide exchange activity are catalyzed by a common domain on the dbl oncogene product. J Biol Chem 269:62–65

    CAS  PubMed  Google Scholar 

  • Hill CS, Wynne J, Treisman R (1995) The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF. Cell 81:1159–1170

    CAS  PubMed  Google Scholar 

  • Jaffe AB, Hall A (2002) Rho GTPases in transformation and metastasis. Adv Cancer Res 84:57–80

    CAS  PubMed  Google Scholar 

  • Kjøller L, Hall A (1999) Signaling to Rho GTPases. Exp Cell Res 253:166–179

    Article  PubMed  Google Scholar 

  • Mackay DJ, Hall A (1998) Rho GTPases. J Biol Chem 273:20685–20688

    Article  CAS  PubMed  Google Scholar 

  • Mao J, Yuan H, Xie W, Wu D (1998) Guanine nucleotide exchange factor GEF115 specifically mediates activation of Rho and serum response factor by the G-protein α subunit Gα13. Proc Natl Acad Sci USA 95:12973–12976

    Article  CAS  PubMed  Google Scholar 

  • Michiels F, Habets GGM, Stam JC, Van der Kammen RA, Collard JG (1995) A role for Rac in Tiam1-induced membrane ruffling and invasion. Nature 375:338–340

    Article  CAS  PubMed  Google Scholar 

  • Ren X, Schwartz MA (2000) Determination of GTP loading on Rho. Methods Enzymol 325:264–272

    CAS  PubMed  Google Scholar 

  • Rümenapp U, Blomquist A, Schwörer G, Schablowski H, Psoma A, Jakobs KH (1999) Rho-specific binding and guanine nucleotide exchange catalysis by KIAA0380, a Dbl family member. FEBS Lett 459:313–318

    Article  PubMed  Google Scholar 

  • Rümenapp U, Freichel-Blomquist A, Wittinghofer B, Jakobs KH, Wieland T (2002) A mammalian Rho-specific guanine-nucleotide exchange factor (p164-RhoGEF) without a pleckstrin homology domain. Biochem J 366:721–728

    PubMed  Google Scholar 

  • Schmidt A, Hall A (2002) Guanine nucleotide exchange factors for Rho GTPases: turning on the switch. Genes Dev 16:1587–1609

    Article  CAS  PubMed  Google Scholar 

  • Souchet M, Portales-Casamar E, Mazurais D, Schmidt S, Leger I, Javré JL, Robert P, Berrebi-Bertrand I, Bril A, Gout B, Debant A, Calmels TPG (2002) Human p63RhoGEF, a novel RhoA-specific guanine nucleotide exchange factor, is localized in cardiac sarcomere. J Cell Sci 115:629–639

    CAS  PubMed  Google Scholar 

  • Stam JC, Collard JG (1999) The DH protein family, exchange factors for Rho-like GTPases. Prog Mol Subcell Biol 22:51–83

    CAS  PubMed  Google Scholar 

  • Zheng Y (2001) Dbl family guanine nucleotide exchange factors. Trends Biochem Sci 26:724–732

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Alexa, A. Hahn, K. Baden and C. Moorkamp for technical assistance. The gifts of various plasmids by Drs. M.M. Chou, R. Fuji, A. Hall, J. Mao, D. Wu and M.A. Schwartz are greatly appreciated. This work was supported by grants from the Deutsche Forschungsgemeinschaft, the Interne Forschungsförderung Essen, the Forschungsfonds of the Fakultät für Klinische Medizin Mannheim and the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Institut für Pharmakologie und Toxikologie, Fakultät für Klinische Medizin Mannheim, Universität Heidelberg, Maybachstrasse 14–16, 68169, Mannheim, Germany

    Susanne Lutz & Thomas Wieland

  2. Institut für Pharmakologie, Universitätsklinikum Essen, Hufelandstrasse 55, 45122, Essen, Germany

    Andrea Freichel-Blomquist, Ulrich Rümenapp, Martina Schmidt & Karl H. Jakobs

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  1. Susanne Lutz

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  2. Andrea Freichel-Blomquist

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  3. Ulrich Rümenapp

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  4. Martina Schmidt

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  5. Karl H. Jakobs

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  6. Thomas Wieland

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Corresponding author

Correspondence to Thomas Wieland.

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Lutz, S., Freichel-Blomquist, A., Rümenapp, U. et al. p63RhoGEF and GEFT are Rho-specific guanine nucleotide exchange factors encoded by the same gene. Naunyn-Schmiedeberg's Arch Pharmacol 369, 540–546 (2004). https://doi.org/10.1007/s00210-004-0926-5

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  • Received: 08 January 2004

  • Accepted: 10 March 2004

  • Published: 07 April 2004

  • Issue Date: May 2004

  • DOI: https://doi.org/10.1007/s00210-004-0926-5

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