J 2014

Disheveled regulates precoupling of heterotrimeric G proteins to Frizzled 6

KILLANDER, Michaela BC, Julian PETERSEN, Kjetil Wessel ANDRESSEN, Sri Ranjani GANJI, Finn Olav LEVY et. al.

Basic information

Original name

Disheveled regulates precoupling of heterotrimeric G proteins to Frizzled 6

Authors

KILLANDER, Michaela BC (752 Sweden), Julian PETERSEN (752 Sweden), Kjetil Wessel ANDRESSEN (578 Norway), Sri Ranjani GANJI (356 India, belonging to the institution), Finn Olav LEVY (578 Norway), Jens SCHUSTER (752 Sweden), Niklas DAHL (752 Sweden), Vítězslav BRYJA (203 Czech Republic, guarantor, belonging to the institution) and Gunnar SCHULTE (276 Germany, belonging to the institution)

Edition

FASEB JOURNAL, 2014, 0892-6638

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 5.043

RIV identification code

RIV/00216224:14310/14:00073587

Organization unit

Faculty of Science

DOI

UT WoS

000335336000031

Keywords in English

GNAI1; GNAQ; GPCR; WNT-5A

Tags

Změněno: 27/4/2015 15:08, Ing. Andrea Mikešková

Abstract

V originále

Frizzleds (FZDs) are classified as G-protein-coupling receptors, but how signals are initiated and specified through heterotrimeric G proteins is unknown. FZD6 regulates convergent extension movements, and its C-terminal Arg511Cys mutation causes nail dysplasia in humans. We investigated the functional relationship between FZD6, Disheveled (DVL), and heterotrimeric G proteins. Live cell imaging combined with fluorescence recovery after photobleaching (FRAP) revealed that inactive human FZD6 precouples to GALPHAi1 and GALPHAq but not to GALPHAoA,GALPHAs, and GALPHA12 proteins. G-protein coupling is measured as a 10-20% reduction in the mobile fraction of fluorescently tagged G proteins on chemical receptor surface cross-linking. The FZD6 Arg511Cys mutation is incapable of G-protein precoupling, even though it still binds DVL. Using both FRAP and Förster resonance energy transfer (FRET) technology, we showed that the FZD6-GALPHAi1 and FZD-GALPHAq complexes dissociate on WNT-5A stimulation. Most important, G-protein precoupling of FZD6 and WNT-5A-induced signaling to extracellular signal-regulated kinase1/2 were impaired by DVL knockdown or overexpression, arguing for a strict dependence of FZD6-G-protein coupling on DVL levels and identifying DVL as a master regulator of FZD/G-protein signaling. In summary, we propose a mechanistic connection between DVL and G proteins integrating WNT, FZD, G-protein, and DVL function.

Links

EE2.3.20.0180, research and development project
Name: Spolupráce mezi Masarykovou univerzitou a Karolinska Institutet, Stockholm na poli biomedicíny
GA13-32990S, research and development project
Name: Posttranslační modifikace receptorů na buněčném povrchu jako určující faktor pro specificitu signálu
Investor: Czech Science Foundation
GA204/09/0498, research and development project
Name: Dynamika proteinů interagujících s Dishevelled a jejich význam pro Wnt signálování
Investor: Czech Science Foundation, Dynamics of proteins interacting with Dishevelled and their importance for Wnt signalling
GD204/09/H058, research and development project
Name: Mezibuněčná signalizace ve vývoji organismu a vzniku onemocnění
Investor: Czech Science Foundation, Intercellular signalling in development and disease
MSM0021622430, plan (intention)
Name: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie
Investor: Ministry of Education, Youth and Sports of the CR, Functional and molecular characteristics of cancer and normal stem cells - identification of targets for novel therapeutics and therapeutic strategies