Splice variants of CK1a and CK1a-like: Comparative analysis of subcellular localization, kinase activity, and function in the Wnt signaling pathway

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F24%3A00603489" target="_blank" >RIV/68081707:_____/24:00603489 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0021925824019082?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0021925824019082?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jbc.2024.107407" target="_blank" >10.1016/j.jbc.2024.107407</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Splice variants of CK1a and CK1a-like: Comparative analysis of subcellular localization, kinase activity, and function in the Wnt signaling pathway

Popis výsledku v původním jazyce

Members of the casein kinase 1 (CK1) family are important regulators of multiple signaling pathways. CK1a is a wellknown negative regulator of the Wnt/b-catenin pathway, which promotes the degradation of b-catenin via its phosphorylation of Ser45. In contrast, the closest paralog of CK1a, CK1a-like, is a poorly characterized kinase of unknown function. In this study, we show that the deletion of CK1a, but not CK1a-like, resulted in a strong activation of the Wnt/b-catenin pathway. Wnt-3a treatment further enhanced the activation, which suggests there are at least two modes, a CK1a-dependent and Wnt-dependent, of b-catenin regulation. Rescue experiments showed that only two out of ten naturally occurring splice CK1a/a-like variants were able to rescue the augmented Wnt/b-catenin signaling caused by CK1a deficiency in cells. Importantly, the ability to phosphorylate b-catenin on Ser45 in the in vitro kinase assay was required but not sufficient for such rescue. Our compound CK1a and GSK3a/b KO models suggest that the additional nonredundant function of CK1a in the Wnt pathway beyond Ser45-b-catenin phosphorylation includes Axin phosphorylation. Finally, we established NanoBRET assays for the three most common CK1a splice variants as well as CK1a-like. Target engagement data revealed comparable potency of known CK1a inhibitors for all CK1a variants but not for CK1a-like. In summary, our work brings important novel insights into the biology of CK1a, including evidence for the lack of redundancy with other CK1 kinases in the negative regulation of the Wnt/b-catenin pathway at the level of b- catenin and Axin.

Název v anglickém jazyce

Splice variants of CK1a and CK1a-like: Comparative analysis of subcellular localization, kinase activity, and function in the Wnt signaling pathway

Popis výsledku anglicky

Members of the casein kinase 1 (CK1) family are important regulators of multiple signaling pathways. CK1a is a wellknown negative regulator of the Wnt/b-catenin pathway, which promotes the degradation of b-catenin via its phosphorylation of Ser45. In contrast, the closest paralog of CK1a, CK1a-like, is a poorly characterized kinase of unknown function. In this study, we show that the deletion of CK1a, but not CK1a-like, resulted in a strong activation of the Wnt/b-catenin pathway. Wnt-3a treatment further enhanced the activation, which suggests there are at least two modes, a CK1a-dependent and Wnt-dependent, of b-catenin regulation. Rescue experiments showed that only two out of ten naturally occurring splice CK1a/a-like variants were able to rescue the augmented Wnt/b-catenin signaling caused by CK1a deficiency in cells. Importantly, the ability to phosphorylate b-catenin on Ser45 in the in vitro kinase assay was required but not sufficient for such rescue. Our compound CK1a and GSK3a/b KO models suggest that the additional nonredundant function of CK1a in the Wnt pathway beyond Ser45-b-catenin phosphorylation includes Axin phosphorylation. Finally, we established NanoBRET assays for the three most common CK1a splice variants as well as CK1a-like. Target engagement data revealed comparable potency of known CK1a inhibitors for all CK1a variants but not for CK1a-like. In summary, our work brings important novel insights into the biology of CK1a, including evidence for the lack of redundancy with other CK1 kinases in the negative regulation of the Wnt/b-catenin pathway at the level of b- catenin and Axin.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Biological Chemistry

ISSN

0021-9258

e-ISSN

1083-351X

Svazek periodika

300

Číslo periodika v rámci svazku

7

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

22

Strana od-do

107407

Kód UT WoS článku

001364964200001

EID výsledku v databázi Scopus

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