Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Functional consequences of Gα13 mutations that disrupt interaction with p115RhoGEF

Oncogene volume 24, pages 2155–2165 (2005)Cite this article

Abstract

The G-protein α subunit, α13, regulates cell growth and differentiation through the monomeric Rho GTPase. α13 activates Rho through direct stimulation of the guanine nucleotide exchange factor p115RhoGEF, which contains a regulator of G-protein signaling homology domain (RH) in its N-terminus. Through its RH domain, p115RhoGEF also functions as a GAP for Gα13. The mechanism for the Gα13/p115RhoGEF interaction is not well understood. Here, we determined specific α13 residues important for its interaction with p115RhoGEF. GST-pulldowns and co-immunoprecipitation assays revealed that individually mutating α13 residues Lys204, Glu229, or Arg232 to opposite charge residues disrupts the interaction of activated α13 with the RH domain of p115RhoGEF or full-length p115RhoGEF. We further demonstrate that mutation of Glu229, and to a lesser extent Lys204 or Arg232, disrupts the ability of activated α13 to induce the recruitment of p115RhoGEF to the plasma membrane (PM) and to activate Rho-mediated serum response element-luciferase gene transcription. Interestingly, an α13 mutant where a conserved Gly was mutated to a Ser (G205S) retained its ability to bind to p115RhoGEF, induce p115RhoGEF recruitment to the PM, and activate Rho-dependent signaling, even though identical Gly to Ser mutations in other α disrupt their interaction with regulator of G-protein signaling (RGS) proteins. These results demonstrate that, whereas several features of a typical α/RGS interaction are preserved in the α13/p115RhoGEF interaction, there are also significant differences.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

Abbreviations

G protein:

guanine nucleotide-binding protein

GPCR:

G protein-coupled receptor

RGS:

regulator of G-protein signaling

RH:

regulator of G-protein signaling homology domain

GEF:

guanine-nucleotide exchange factor

LARG:

leukemia-associated RhoGEF

GAP:

GTPase activating protein

SRE:

serum response element

PM:

plasma membrane

GRK:

GPCR kinase

GFP:

green fluorescent protein

HA:

hemagglutinin

Ni-NTA:

nickel-nitrilotriacetic acid

References

  • Althoefer H, Eversole-Cire P and Simon MI . (1997). J. Biol. Chem., 272, 24380–24386.

  • Berestetskaya YV, Faure MP, Ichijo H and Voyno-Yasenetskaya TA . (1998). J. Biol. Chem., 273, 27816–27823.

  • Bhattacharyya R and Wedegaertner PB . (2000). J. Biol. Chem., 275, 14992–14999.

  • Bhattacharyya R and Wedegaertner PB . (2003a). Biochem. J., 371, 709–720.

  • Bhattacharyya R and Wedegaertner PB . (2003b). FEBS Lett., 540, 211–216.

  • Buhl AM, Johnson NL, Dhanasekaran N and Johnson GL . (1995). J. Biol. Chem., 270, 24631–24634.

  • Chen Z, Singer WD, Wells CD, Sprang SR and Sternweis PC . (2003). J. Biol. Chem., 278, 9912–9919.

  • Chen Z, Wells CD, Sternweis PC and Sprang SR . (2001). Nat. Struct. Biol., 8, 805–809.

  • Coleman DE, Berghuis AM, Lee E, Linder ME, Gilman AG and Sprang SR . (1994). Science, 265, 1405–1412.

  • DiBello PR, Garrison TR, Apanovitch DM, Hoffman G, Shuey DJ, Mason K, Cockett MI and Dohlman HG . (1998). J. Biol. Chem., 273, 5780–5784.

  • Evanko DS, Thiyagarajan MM and Wedegaertner PB . (2000). J. Biol. Chem., 275, 1327–1336.

  • Fleming IN, Elliott CM and Exton JH . (1996). J. Biol. Chem., 271, 33067–33073.

  • Fukuhara S, Murga C, Zohar M, Igishi T and Gutkind JS . (1999). J. Biol. Chem., 274, 5868–5879.

  • Gohla A, Offermanns S, Wilkie TM and Schultz G . (1999). J. Biol. Chem., 274, 17901–17907.

  • Hamm HE . (1998). J. Biol. Chem., 273, 669–672.

  • Hart MJ, Jiang X, Kozasa T, Roscoe W, Singer WD, Gilman AG, Sternweis PC and Bollag G . (1998). Science, 280, 2112–2114.

  • Hollinger S and Hepler JR . (2002). Pharmacol. Rev., 54, 527–559.

  • Jiang H, Wu D and Simon MI . (1993). FEBS Lett., 330, 319–322.

  • Katoh H, Aoki J, Yamaguchi Y, Kitano Y, Ichikawa A and Negishi M . (1998). J. Biol. Chem., 273, 28700–28707.

  • Kozasa T, Jiang X, Hart MJ, Sternweis PM, Singer WD, Gilman AG, Bollag G and Sternweis PC . (1998). Science, 280, 2109–2111.

  • Kranenburg O, Poland M, Gebbink M, Oomen L and Moolenaar WH . (1997). J. Cell Sci., 110, 2417–2427.

  • Kranenburg O, Poland M, van Horck FP, Drechsel D, Hall A and Moolenaar WH . (1999). Mol. Biol. Cell, 10, 1851–1857.

  • Lan KL, Sarvazyan NA, Taussig R, Mackenzie RG, DiBello PR, Dohlman HG and Neubig RR . (1998). J. Biol. Chem., 273, 12794–12797.

  • Mao J, Xie W, Yuan H, Simon MI, Mano H and Wu D . (1998a). EMBO J., 17, 5638–5646.

  • Mao J, Yuan H, Xie W, Simon MI and Wu D . (1998b). J. Biol. Chem., 273, 27118–27123.

  • Mao J, Yuan H, Xie W and Wu D . (1998c). Proc. Natl. Acad. Sci. USA, 95, 12973–12976.

  • Meigs TE, Fedor-Chaiken M, Kaplan DD, Brackenbury R and Casey PJ . (2002). J. Biol. Chem., 277, 24594–24600.

  • Meigs TE, Fields TA, McKee DD and Casey PJ . (2001). Proc. Natl. Acad. Sci. USA, 98, 519–524.

  • Nakamura S, Kreutz B, Tanabe S, Suzuki N and Kozasa T . (2004). Mol. Pharmacol., 66, 1029–1034.

  • Neubig RR . (1994). FASEB J., 8, 939–946.

  • Offermanns S . (2000). Rev. Physiol. Biochem. Pharmacol., 140, 63–133.

  • Parks S and Wieschaus E . (1991). Cell, 64, 447–458.

  • Prasad MV, Dermott JM, Heasley LE, Johnson GL and Dhanasekaran N . (1995). J. Biol. Chem., 270, 18655–18659.

  • Shi CS, Sinnarajah S, Cho H, Kozasa T and Kehrl JH . (2000). J. Biol. Chem., 275, 24470–24476.

  • Sterne-Marr R, Tesmer JJ, Day PW, Stracquatanio RP, Cilente JA, O'Connor KE, Pronin AN, Benovic JL and Wedegaertner PB . (2003). J. Biol. Chem., 278, 6050–6058.

  • Symons M . (1996). Trends Biochem. Sci., 21, 178–181.

  • Takida S and Wedegaertner PB . (2003). J. Biol. Chem., 278, 17284–17290.

  • Tesmer JJ, Berman DM, Gilman AG and Sprang SR . (1997). Cell, 89, 251–261.

  • Vaiskunaite R, Adarichev V, Furthmayr H, Kozasa T, Gudkov A and Voyno-Yasenetskaya TA . (2000). J. Biol. Chem., 275, 26206–26212.

  • Voyno-Yasenetskaya T, Conklin BR, Gilbert RL, Hooley R, Bourne HR and Barber DL . (1994a). J. Biol. Chem., 269, 4721–4724.

  • Voyno-Yasenetskaya TA, Faure MP, Ahn NG and Bourne HR . (1996). J. Biol. Chem., 271, 21081–21087.

  • Voyno-Yasenetskaya TA, Pace AM and Bourne HR . (1994b). Oncogene, 9, 2559–2565.

  • Wells C, Jiang X, Gutowski S and Sternweis PC . (2002a). Methods Enzymol., 345, 371–382.

  • Wells CD, Gutowski S, Bollag G and Sternweis PC . (2001). J. Biol. Chem., 276, 28897–28905.

  • Wells CD, Liu MY, Jackson M, Gutowski S, Sternweis PM, Rothstein JD, Kozasa T and Sternweis PC . (2002b). J. Biol. Chem., 277, 1174–1181.

  • Xu N, Bradley L, Ambdukar I and Gutkind JS . (1993). Proc. Natl. Acad. Sci. USA, 90, 6741–6745.

  • Yamaguchi Y, Katoh H, Mori K and Negishi M . (2002). Curr. Biol., 12, 1353–1358.

  • Yamaguchi Y, Katoh H and Negishi M . (2003). J. Biol. Chem., 278, 14936–14939.

Download references

Acknowledgements

This work was supported by grant GM62884 (PW) from the NIH. PW is an Established Investigator of the American Heart Association.

Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, 233 S. 10th Street, 839 BLSB, Philadelphia, PA, 19107, USA

    Elda Grabocka & Philip B Wedegaertner

Authors
  1. Elda Grabocka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Philip B Wedegaertner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Philip B Wedegaertner.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grabocka, E., Wedegaertner, P. Functional consequences of Gα13 mutations that disrupt interaction with p115RhoGEF. Oncogene 24, 2155–2165 (2005). https://doi.org/10.1038/sj.onc.1208414

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1208414

Keywords

This article is cited by

Search

Advanced search

Quick links