Regulator of G protein signaling (RGS) proteins function as GTPase-activating proteins for the α-subunit of heterotrimeric G proteins. The function of certain RGS proteins is negatively regulated by 14-3-3 proteins, a family of highly conserved regulatory molecules expressed in all eukaryotes. In this study, we provide a structural mechanism for 14-3-3-dependent inhibition of RGS3-Gα interaction. We have used small angle x-ray scattering, hydrogen/deuterium exchange kinetics, and Förster resonance energy transfer measurements to determine the low-resolution solution structure of the 14-3-3ζ·RGS3 complex. The structure shows the RGS domain of RGS3 bound to the 14-3-3ζ dimer in an as-yet-unrecognized manner interacting with less conserved regions on the outer surface of the 14-3-3 dimer outside its central channel. Our results suggest that the 14-3-3 protein binding affects the structure of the Gα interaction portion of RGS3 as well as sterically blocks the interaction between the RGS domain and the Gα subunit of heterotrimeric G proteins.

Department of Physical and Macromolecular Chemistry Charles University Prague 12843 Prague Czech Republic

Zobrazit více v PubMed

Lee R. H., Whelan J. P., Lolley R. N., McGinnis J. F. (1988) Exp. Eye Res. 46, 829–840 PubMed

Willardson B. M., Wilkins J. F., Yoshida T., Bitensky M. W. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 1475–1479 PubMed PMC

Berman D. M., Wilkie T. M., Gilman A. G. (1996) Cell 86, 445–452 PubMed

Watson N., Linder M. E., Druey K. M., Kehrl J. H., Blumer K. J. (1996) Nature 383, 172–175 PubMed

Hepler J. R., Berman D. M., Gilman A. G., Kozasa T. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 428–432 PubMed PMC

Tesmer J. J., Berman D. M., Gilman A. G., Sprang S. R. (1997) Cell 89, 251–261 PubMed

De Vries L., Gist Farquhar M. (1999) Trends Cell Biol. 9, 138–144 PubMed

Abramow-Newerly M., Roy A. A., Nunn C., Chidiac P. (2006) Cell. Signal. 18, 579–591 PubMed

Benzing T., Yaffe M. B., Arnould T., Sellin L., Schermer B., Schilling B., Schreiber R., Kunzelmann K., Leparc G. G., Kim E., Walz G. (2000) J. Biol. Chem. 275, 28167–28172 PubMed

Benzing T., Köttgen M., Johnson M., Schermer B., Zentgraf H., Walz G., Kim E. (2002) J. Biol. Chem. 277, 32954–32962 PubMed

Niu J., Scheschonka A., Druey K. M., Davis A., Reed E., Kolenko V., Bodnar R., Voyno-Yasenetskaya T., Du X., Kehrl J., Dulin N. O. (2002) Biochem. J. 365, 677–684 PubMed PMC

Ward R. J., Milligan G. (2005) Mol. Pharmacol. 68, 1821–1830 PubMed

Abramow-Newerly M., Ming H., Chidiac P. (2006) Cell. Signal. 18, 2209–2222 PubMed

Fu H., Subramanian R. R., Masters S. C. (2000) Annu. Rev. Pharmacol. Toxicol. 40, 617–647 PubMed

Bridges D., Moorhead G. B. (2004) Sci. STKE 2004, re10. PubMed

Mackintosh C. (2004) Biochem. J. 381, 329–342 PubMed PMC

Aitken A. (2006) Semin. Cancer Biol. 16, 162–172 PubMed

Xiao B., Smerdon S. J., Jones D. H., Dodson G. G., Soneji Y., Aitken A., Gamblin S. J. (1995) Nature 376, 188–191 PubMed

Liu D., Bienkowska J., Petosa C., Collier R. J., Fu H., Liddington R. (1995) Nature 376, 191–194 PubMed

Muslin A. J., Tanner J. W., Allen P. M., Shaw A. S. (1996) Cell 84, 889–897 PubMed

Rittinger K., Budman J., Xu J., Volinia S., Cantley L. C., Smerdon S. J., Gamblin S. J., Yaffe M. B. (1999) Mol. Cell 4, 153–166 PubMed

Yaffe M. B., Rittinger K., Volinia S., Caron P. R., Aitken A., Leffers H., Gamblin S. J., Smerdon S. J., Cantley L. C. (1997) Cell 91, 961–971 PubMed

Petosa C., Masters S. C., Bankston L. A., Pohl J., Wang B., Fu H., Liddington R. C. (1998) J. Biol. Chem. 273, 16305–16310 PubMed

Obsil T., Ghirlando R., Klein D. C., Ganguly S., Dyda F. (2001) Cell 105, 257–267 PubMed

Ottmann C., Marco S., Jaspert N., Marcon C., Schauer N., Weyand M., Vandermeeren C., Duby G., Boutry M., Wittinghofer A., Rigaud J. L., Oecking C. (2007) Mol. Cell 25, 427–440 PubMed

Silhan J., Vacha P., Strnadova P., Vecer J., Herman P., Sulc M., Teisinger J., Obsilova V., Obsil T. (2009) J. Biol. Chem. 284, 19349–19360 PubMed PMC

Rezabkova L., Boura E., Herman P., Vecer J., Bourova L., Sulc M., Svoboda P., Obsilova V., Obsil T. (2010) J. Struct. Biol. 170, 451–461 PubMed

Silhan J., Obsilova V., Vecer J., Herman P., Sulc M., Teisinger J., Obsil T. (2004) J. Biol. Chem. 279, 49113–49119 PubMed

Boura E., Silhan J., Herman P., Vecer J., Sulc M., Teisinger J., Obsilova V., Obsil T. (2007) J. Biol. Chem. 282, 8265–8275 PubMed

Woodcock J. M., Murphy J., Stomski F. C., Berndt M. C., Lopez A. F. (2003) J. Biol. Chem. 278, 36323–36327 PubMed

Houtman J. C., Brown P. H., Bowden B., Yamaguchi H., Appella E., Samelson L. E., Schuck P. (2007) Protein Sci. 16, 30–42 PubMed PMC

Schuck P. (2000) Biophys. J. 78, 1606–1619 PubMed PMC

Roessle M. W., Klaering R., Ristau U., Robrahn B., Jahn D., Gehrmann T., Konarev P., Round A., Fiedler S., Hermes C., Svergun D. (2007) J. Appl. Crystallogr. 40, S190–S194

Guinier A. (1939) Ann. Phys. (Paris) 12, 161–237

Svergun D. I. (1992) J. Appl. Crystallogr. 25, 495–503

Porod G. (1982) in Small-angle X-ray Scattering (Glatter O., Kratky O. eds) pp. 17–51, Academic Press, London

Franke D., Svergun D. I. (2009) J. Appl. Crystallogr. 42, 342–346 PubMed PMC

Volkov V. V., Svergun D. I. (2003) J. Appl. Crystallogr. 36, 860–864

Strohalm M., Kavan D., Novák P., Volný M., Havlícek V. (2010) Anal. Chem. 82, 4648–4651 PubMed

Veisova D., Rezabkova L., Stepanek M., Novotna P., Herman P., Vecer J., Obsil T., Obsilova V. (2010) Biochemistry 49, 3853–3861 PubMed

Obsilova V., Herman P., Vecer J., Sulc M., Teisinger J., Obsil T. (2004) J. Biol. Chem. 279, 4531–4540 PubMed

Vecer J., Herman P. (2011) J. Fluoresc. 21, 873–881 PubMed

Melhuish W. H. (1961) J. Phys. Chem. 65, 229–235

Yang X., Lee W. H., Sobott F., Papagrigoriou E., Robinson C. V., Grossmann J. G., Sundström M., Doyle D. A., Elkins J. M. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 17237–17242 PubMed PMC

Engen J. R. (2003) Analyst 128, 623–628 PubMed

Marcoux J., Man P., Petit-Haertlein I., Vivès C., Forest E., Fieschi F. (2010) J. Biol. Chem. 285, 28980–28990 PubMed PMC

Lakowicz J. R. (1999) Principles of Fluorescence Spectroscopy, Second Ed., pp. 321–346, Kluwer Academic/Plenum Publishers, New York

Davezac N., Baldin V., Gabrielli B., Forrest A., Theis-Febvre N., Yashida M., Ducommun B. (2000) Oncogene 19, 2179–2185 PubMed

Margolis S. S., Kornbluth S. (2004) Cell Cycle 3, 425–428 PubMed

McKinsey T. A., Zhang C. L., Olson E. N. (2001) Mol. Cell. Biol. 21, 6312–6321 PubMed PMC

Yang J., Winkler K., Yoshida M., Kornbluth S. (1999) EMBO J. 18, 2174–2183 PubMed PMC

Brunet A., Bonni A., Zigmond M. J., Lin M. Z., Juo P., Hu L. S., Anderson M. J., Arden K. C., Blenis J., Greenberg M. E. (1999) Cell 96, 857–868 PubMed

Brownawell A. M., Kops G. J., Macara I. G., Burgering B. M. (2001) Mol. Cell. Biol. 21, 3534–3546 PubMed PMC

Obsilova V., Vecer J., Herman P., Pabianova A., Sulc M., Teisinger J., Boura E., Obsil T. (2005) Biochemistry 44, 11608–11617 PubMed

Benzinger A., Popowicz G. M., Joy J. K., Majumdar S., Holak T. A., Hermeking H. (2005) Cell Res. 15, 219–227 PubMed

Wilker E. W., Grant R. A., Artim S. C., Yaffe M. B. (2005) J. Biol. Chem. 280, 18891–18898 PubMed

Gardino A. K., Smerdon S. J., Yaffe M. B. (2006) Semin. Cancer Biol. 16, 173–182 PubMed

Bustos D. M., Iglesias A. A. (2006) Proteins 63, 35–42 PubMed

The interaction of the mitochondrial protein importer TOMM34 with HSP70 is regulated by TOMM34 phosphorylation and binding to 14-3-3 adaptors

Coordination and redox state-dependent structural changes of the heme-based oxygen sensor AfGcHK associated with intraprotein signal transduction

Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1

Novel Entropically Driven Conformation-specific Interactions with Tomm34 Protein Modulate Hsp70 Protein Folding and ATPase Activities

Structural Characterization of Phosducin and Its Complex with the 14-3-3 Protein

Role of the EF-hand-like motif in the 14-3-3 protein-mediated activation of yeast neutral trehalase Nth1

The assembly and intermolecular properties of the Hsp70-Tomm34-Hsp90 molecular chaperone complex

Structural modulation of phosducin by phosphorylation and 14-3-3 protein binding