The N-Terminal Part of the Dishevelled DEP Domain Is Required
for Wnt/beta-Catenin Signaling in Mammalian Cells

J 2017

The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells

PACLÍKOVÁ, Petra, Ondřej BERNATÍK, Tomasz Witold RADASZKIEWICZ a Vítězslav BRYJA

Základní údaje

Originální název

The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells

Autoři

PACLÍKOVÁ, Petra (203 Česká republika, domácí), Ondřej BERNATÍK (203 Česká republika, domácí), Tomasz Witold RADASZKIEWICZ (616 Polsko, domácí) a Vítězslav BRYJA (203 Česká republika, garant, domácí)

Vydání

Molecular and cellular biology, WASHINGTON, AMER SOC MICROBIOLOGY, 2017, 0270-7306

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10601 Cell biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.813

Kód RIV

RIV/00216224:14310/17:00094935

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1128/MCB.00145-17

UT WoS

000408425300005

Klíčová slova anglicky

CRISPR/Cas; DEP domain; Dishevelled; Wnt/beta-catenin signaling; Wnt3a

Štítky

NZ, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 12. 4. 2018 14:20, Ing. Nicole Zrilić

Anotace

V originále

Dishevelled (DVL) proteins are key mediators of the Wnt/beta-catenin signaling pathway. All DVL proteins contain three conserved domains: DIX, PDZ, and DEP. There is a consensus in the field that the DIX domain is critical for Wnt/beta-catenin signaling, but contradictory evidence regarding the function of the DEP domain exists. It has been difficult, until recently, to test the importance of the DEP domain rigorously because of the interference with endogenous DVL, expressed in all Wnt-responsive cell lines. In this study, we took advantage of DVL knockout (DVL1/DVL2/DVL3 triple knockout) cells fully deficient in Wnt3a-induced signaling events and performed a series of rescue experiments. Using these complementation assays, we analyzed the role of individual DVL isoforms. Further domain mapping of DVL1 showed that both the DVL1 DEP domain and especially its N-terminal region are required and sufficient for Wnt3a-induced phosphorylation of LRP6 and TopFlash reporter activation. On the contrary, multiple DEP domain mutants deficient in the planar cell polarity (PCP) pathway could fully rescue the Wnt3a response. This study provides conclusive evidence that the DVL DEP domain is essential for Wnt/beta-catenin signaling in mammalian cells and establishes an experimental system suitable for further functional testing of DVL.

Návaznosti

GA15-21789S, projekt VaV

Název: Dishevelled - identifikace základních koordinát

Investor: Grantová agentura ČR, Dishevelled - identifikace základních koordinát

GA17-16680S, projekt VaV

Název: Nové postupy pro určení aktivity dráhy planární buněčné polarity (PCP)

Investor: Grantová agentura ČR, Nové postupy pro určení aktivity dráhy planární buněčné polarity (PCP)

MUNI/G/1100/2016, interní kód MU

Název: Computational chemistry for Wnt signaling pathway

Investor: Masarykova univerzita, Computational chemistry for Wnt signaling pathway, INTERDISCIPLINARY - Mezioborové výzkumné projekty

Citovat

PACLÍKOVÁ, Petra, Ondřej BERNATÍK, Tomasz Witold RADASZKIEWICZ a Vítězslav BRYJA. The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells. Molecular and cellular biology. WASHINGTON: AMER SOC MICROBIOLOGY, 2017, roč. 37, č. 18, s. nestránkováno, 16 s. ISSN 0270-7306. Dostupné z: https://dx.doi.org/10.1128/MCB.00145-17.

@article{1389333,
   author = {Paclíková, Petra and Bernatík, Ondřej and Radaszkiewicz, Tomasz Witold and Bryja, Vítězslav},
   article_location = {WASHINGTON},
   article_number = {18},
   doi = {http://dx.doi.org/10.1128/MCB.00145-17},
   keywords = {CRISPR/Cas; DEP domain; Dishevelled; Wnt/beta-catenin signaling; Wnt3a},
   language = {eng},
   issn = {0270-7306},
   journal = {Molecular and cellular biology},
   title = {The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells},
   volume = {37},
   year = {2017}
}

TY  - JOUR
ID  - 1389333
AU  - Paclíková, Petra - Bernatík, Ondřej - Radaszkiewicz, Tomasz Witold - Bryja, Vítězslav
PY  - 2017
TI  - The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells
JF  - Molecular and cellular biology
VL  - 37
IS  - 18
SP  - nestránkováno
EP  - nestránkováno
PB  - AMER SOC MICROBIOLOGY
SN  - 02707306
KW  - CRISPR/Cas
KW  - DEP domain
KW  - Dishevelled
KW  - Wnt/beta-catenin signaling
KW  - Wnt3a
N2  - Dishevelled (DVL) proteins are key mediators of the Wnt/beta-catenin signaling pathway. All DVL proteins contain three conserved domains: DIX, PDZ, and DEP. There is a consensus in the field that the DIX domain is critical for Wnt/beta-catenin signaling, but contradictory evidence regarding the function of the DEP domain exists. It has been difficult, until recently, to test the importance of the DEP domain rigorously because of the interference with endogenous DVL, expressed in all Wnt-responsive cell lines. In this study, we took advantage of DVL knockout (DVL1/DVL2/DVL3 triple knockout) cells fully deficient in Wnt3a-induced signaling events and performed a series of rescue experiments. Using these complementation assays, we analyzed the role of individual DVL isoforms. Further domain mapping of DVL1 showed that both the DVL1 DEP domain and especially its N-terminal region are required and sufficient for Wnt3a-induced phosphorylation of LRP6 and TopFlash reporter activation. On the contrary, multiple DEP domain mutants deficient in the planar cell polarity (PCP) pathway could fully rescue the Wnt3a response. This study provides conclusive evidence that the DVL DEP domain is essential for Wnt/beta-catenin signaling in mammalian cells and establishes an experimental system suitable for further functional testing of DVL.
ER  -

PACLÍKOVÁ, Petra, Ondřej BERNATÍK, Tomasz Witold RADASZKIEWICZ a Vítězslav BRYJA. The N-Terminal Part of the Dishevelled DEP Domain Is Required for Wnt/beta-Catenin Signaling in Mammalian Cells. \textit{Molecular and cellular biology}. WASHINGTON: AMER SOC MICROBIOLOGY, 2017, roč.~37, č.~18, s.~nestránkováno, 16 s. ISSN~0270-7306. Dostupné z: https://dx.doi.org/10.1128/MCB.00145-17.

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