Roles of individual human Dishevelled paralogs in the Wnt
signalling pathways

J 2021

Roles of individual human Dishevelled paralogs in the Wnt signalling pathways

PACLÍKOVÁ, Petra, Tomasz Witold RADASZKIEWICZ, David POTĚŠIL, Jakub HARNOŠ, Zbyněk ZDRÁHAL et. al.

Basic information

Original name

Roles of individual human Dishevelled paralogs in the Wnt signalling pathways

Authors

PACLÍKOVÁ, Petra (203 Czech Republic, belonging to the institution), Tomasz Witold RADASZKIEWICZ (616 Poland, belonging to the institution), David POTĚŠIL (203 Czech Republic, belonging to the institution), Jakub HARNOŠ (203 Czech Republic, belonging to the institution), Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution) and Vítězslav BRYJA (203 Czech Republic, guarantor, belonging to the institution)

Edition

Cellular Signalling, Elsevier, 2021, 0898-6568

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 4.850

RIV identification code

RIV/00216224:14310/21:00124350

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.cellsig.2021.110058

UT WoS

000685349100004

Keywords in English

Dishevelled 1; 2; 3; Wnt signalling; ROR1; Axin1; CRISPR; Cas9

Abstract

V originále

Dishevelled (DVL) proteins are key mediators of most Wnt pathways. In all vertebrates, three DVL paralogs are present (DVL1, DVL2 and DVL3) but it is poorly defined to what extent they are functionally redundant. Here, we generated T-REx HEK 293 cells with only one DVL paralog (i.e., DVL1-only, DVL2-only, and DVL3-only) and compared their response to Wnt-3a and Wnt-5a ligands with wild type and DVL triple knockout cells. We show that DVL is essential, in addition to the previously shown Wnt-3a-induced phosphorylation of LRP6 and transcriptional activation of TCF/LEF-dependent reporter, also for Wnt-3a-induced degradation of AXIN1 and Wnt5a-induced phosphorylation of ROR1. We have quantified the molar ratios of DVL1:DVL2:DVL3 in our model to be approximately 4:80:16. Interestingly, DVL-only cells do not compensate for the lack of other paralogs and are still fully functional in all analyzed readouts with the exception of Wnt-3a-induced transcription assessed by TopFlash assay. In this assay, the DVL1-only cell line was the most potent; on the contrary, the DVL3-only cell line exhibited only the negligible capacity to mediate Wnt signals. Using a novel model system - complementation assays in T-REx HEK 293 with amplified Wnt signal response (RNF43/ZNRF3/DVL1/DVL2/DVL3 penta KO cells) we demonstrate that it is not the total amount of DVL but ratio of individual paralogs what decides the signal strength. In sum, this study contributes to our better understanding of the role of individual human DVL paralogs in the Wnt pathway.

Links

EF16_025/0007381, research and development project

Name: Preklinická progrese nových organických sloučenin s cílenou biologickou aktivitou

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

90127, large research infrastructures

Name: CIISB II

Citovat

PACLÍKOVÁ, Petra, Tomasz Witold RADASZKIEWICZ, David POTĚŠIL, Jakub HARNOŠ, Zbyněk ZDRÁHAL and Vítězslav BRYJA. Roles of individual human Dishevelled paralogs in the Wnt signalling pathways. Cellular Signalling. Elsevier, 2021, vol. 85, September, p. 110058-110068. ISSN 0898-6568. Available from: https://dx.doi.org/10.1016/j.cellsig.2021.110058.

@article{1838561,
   author = {Paclíková, Petra and Radaszkiewicz, Tomasz Witold and Potěšil, David and Harnoš, Jakub and Zdráhal, Zbyněk and Bryja, Vítězslav},
   article_number = {September},
   doi = {http://dx.doi.org/10.1016/j.cellsig.2021.110058},
   keywords = {Dishevelled 1; 2; 3; Wnt signalling; ROR1; Axin1; CRISPR; Cas9},
   language = {eng},
   issn = {0898-6568},
   journal = {Cellular Signalling},
   title = {Roles of individual human Dishevelled paralogs in the Wnt signalling pathways},
   url = {https://www.sciencedirect.com/science/article/pii/S0898656821001479#!},
   volume = {85},
   year = {2021}
}

TY  - JOUR
ID  - 1838561
AU  - Paclíková, Petra - Radaszkiewicz, Tomasz Witold - Potěšil, David - Harnoš, Jakub - Zdráhal, Zbyněk - Bryja, Vítězslav
PY  - 2021
TI  - Roles of individual human Dishevelled paralogs in the Wnt signalling pathways
JF  - Cellular Signalling
VL  - 85
IS  - September
SP  - 110058
EP  - 110058
PB  - Elsevier
SN  - 08986568
KW  - Dishevelled 1
KW  - 2
KW  - 3
KW  - Wnt signalling
KW  - ROR1
KW  - Axin1
KW  - CRISPR
KW  - Cas9
UR  - https://www.sciencedirect.com/science/article/pii/S0898656821001479#!
N2  - Dishevelled (DVL) proteins are key mediators of most Wnt pathways. In all vertebrates, three DVL paralogs are present (DVL1, DVL2 and DVL3) but it is poorly defined to what extent they are functionally redundant. Here, we generated T-REx HEK 293 cells with only one DVL paralog (i.e., DVL1-only, DVL2-only, and DVL3-only) and compared their response to Wnt-3a and Wnt-5a ligands with wild type and DVL triple knockout cells. We show that DVL is essential, in addition to the previously shown Wnt-3a-induced phosphorylation of LRP6 and transcriptional activation of TCF/LEF-dependent reporter, also for Wnt-3a-induced degradation of AXIN1 and Wnt5a-induced phosphorylation of ROR1. We have quantified the molar ratios of DVL1:DVL2:DVL3 in our model to be approximately 4:80:16. Interestingly, DVL-only cells do not compensate for the lack of other paralogs and are still fully functional in all analyzed readouts with the exception of Wnt-3a-induced transcription assessed by TopFlash assay. In this assay, the DVL1-only cell line was the most potent; on the contrary, the DVL3-only cell line exhibited only the negligible capacity to mediate Wnt signals. Using a novel model system - complementation assays in T-REx HEK 293 with amplified Wnt signal response (RNF43/ZNRF3/DVL1/DVL2/DVL3 penta KO cells) we demonstrate that it is not the total amount of DVL but ratio of individual paralogs what decides the signal strength. In sum, this study contributes to our better understanding of the role of individual human DVL paralogs in the Wnt pathway.
ER  -

PACLÍKOVÁ, Petra, Tomasz Witold RADASZKIEWICZ, David POTĚŠIL, Jakub HARNOŠ, Zbyněk ZDRÁHAL and Vítězslav BRYJA. Roles of individual human Dishevelled paralogs in the Wnt signalling pathways. \textit{Cellular Signalling}. Elsevier, 2021, vol.~85, September, p.~110058-110068. ISSN~0898-6568. Available from: https://dx.doi.org/10.1016/j.cellsig.2021.110058.

Detailed Information on Publication Record