Protease associated domain of RNF43 is not necessary for the
suppression of Wnt/beta-catenin signaling in human cells

J 2020

Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells

RADASZKIEWICZ, Tomasz Witold a Vítězslav BRYJA

Základní údaje

Originální název

Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells

Autoři

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

Vydání

Cell Communication and Signaling, London, BMC, 2020, 1478-811X

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10601 Cell biology

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.712

Kód RIV

RIV/00216224:14310/20:00114371

Organizační jednotka

Přírodovědecká fakulta

DOI

http://dx.doi.org/10.1186/s12964-020-00559-0

UT WoS

000542292400001

Klíčová slova anglicky

RNF43; Protease associated domain; Wnt signaling; LRP6; RSPO1; Dishevelled

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 19. 10. 2020 16:59, Mgr. Marie Šípková, DiS.

Anotace

V originále

BackgroundRNF43 and its homolog ZNRF3 are transmembrane E3 ubiquitin ligases frequently mutated in many human cancer types. Their main role relays on the inhibition of canonical Wnt signaling by the negative regulation of frizzled receptors and LRP5/6 co-receptors levels at the plasma membrane. Intracellular RING domains of RNF43/ZNRF3 mediate the key enzymatic activity of these proteins, but the function of the extracellular Protease Associated (PA) fold in the inhibition of Wnt/beta-catenin pathway is controversial up-to date, apart from the interaction with secreted antagonists R-spondin family proteins shown by the crystallographic studies.MethodsIn our research we utilised cell-based approaches to study the role of RNF43 lacking PA domain in the canonical Wnt signalling pathway transduction. We developed controlled overexpression (TetON) and CRISPR/Cas9 mediated knock-out models in human cells.ResultsRNF43 Delta PA mutant activity impedes canonical Wnt pathway, as manifested by the reduced phosphorylation of LRP6, DVL2 and DVL3 and by the decreased beta-catenin-dependent gene expression. Finally, rescue experiments in the CRISPR/Cas9 derived RNF43/ZNRF3 double knock-out cell lines showed that RNF Delta PA overexpression is enough to inhibit activation of LRP6 and beta-catenin activity as shown by the Western blot and Top flash dual luciferase assays. Moreover, RNF43 variant without PA domain was not sensitive to the R-spondin1 treatment.ConclusionTaken together, our results help to understand better the mode of RNF43 tumor suppressor action and solve some discrepancies present in the field.

Návaznosti

GX19-28347X, projekt VaV

Název: Molekulární a funkční analýza biologie kasein kinázy 1

Investor: Grantová agentura ČR, Molekulární a funkční analýza biologie kasein kinázy 1

Citovat

RADASZKIEWICZ, Tomasz Witold a Vítězslav BRYJA. Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells. Cell Communication and Signaling. London: BMC, 2020, roč. 18, č. 1, s. 1-12. ISSN 1478-811X. Dostupné z: https://dx.doi.org/10.1186/s12964-020-00559-0.

@article{1686033,
   author = {Radaszkiewicz, Tomasz Witold and Bryja, Vítězslav},
   article_location = {London},
   article_number = {1},
   doi = {http://dx.doi.org/10.1186/s12964-020-00559-0},
   keywords = {RNF43; Protease associated domain; Wnt signaling; LRP6; RSPO1; Dishevelled},
   language = {eng},
   issn = {1478-811X},
   journal = {Cell Communication and Signaling},
   title = {Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells},
   url = {https://doi.org/10.1186/s12964-020-00559-0},
   volume = {18},
   year = {2020}
}

TY  - JOUR
ID  - 1686033
AU  - Radaszkiewicz, Tomasz Witold - Bryja, Vítězslav
PY  - 2020
TI  - Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells
JF  - Cell Communication and Signaling
VL  - 18
IS  - 1
SP  - 1-12
EP  - 1-12
PB  - BMC
SN  - 1478811X
KW  - RNF43
KW  - Protease associated domain
KW  - Wnt signaling
KW  - LRP6
KW  - RSPO1
KW  - Dishevelled
UR  - https://doi.org/10.1186/s12964-020-00559-0
L2  - https://doi.org/10.1186/s12964-020-00559-0
N2  - BackgroundRNF43 and its homolog ZNRF3 are transmembrane E3 ubiquitin ligases frequently mutated in many human cancer types. Their main role relays on the inhibition of canonical Wnt signaling by the negative regulation of frizzled receptors and LRP5/6 co-receptors levels at the plasma membrane. Intracellular RING domains of RNF43/ZNRF3 mediate the key enzymatic activity of these proteins, but the function of the extracellular Protease Associated (PA) fold in the inhibition of Wnt/beta-catenin pathway is controversial up-to date, apart from the interaction with secreted antagonists R-spondin family proteins shown by the crystallographic studies.MethodsIn our research we utilised cell-based approaches to study the role of RNF43 lacking PA domain in the canonical Wnt signalling pathway transduction. We developed controlled overexpression (TetON) and CRISPR/Cas9 mediated knock-out models in human cells.ResultsRNF43 Delta PA mutant activity impedes canonical Wnt pathway, as manifested by the reduced phosphorylation of LRP6, DVL2 and DVL3 and by the decreased beta-catenin-dependent gene expression. Finally, rescue experiments in the CRISPR/Cas9 derived RNF43/ZNRF3 double knock-out cell lines showed that RNF Delta PA overexpression is enough to inhibit activation of LRP6 and beta-catenin activity as shown by the Western blot and Top flash dual luciferase assays. Moreover, RNF43 variant without PA domain was not sensitive to the R-spondin1 treatment.ConclusionTaken together, our results help to understand better the mode of RNF43 tumor suppressor action and solve some discrepancies present in the field.
ER  -

RADASZKIEWICZ, Tomasz Witold a Vítězslav BRYJA. Protease associated domain of RNF43 is not necessary for the suppression of Wnt/beta-catenin signaling in human cells. \textit{Cell Communication and Signaling}. London: BMC, 2020, roč.~18, č.~1, s.~1-12. ISSN~1478-811X. Dostupné z: https://dx.doi.org/10.1186/s12964-020-00559-0.

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