NLK inhibits the transcription activity of Smad4 independent of its kinase activity
10.7644/j.issn.1674-9960.2014.03.007
- VernacularTitle:NLK以激酶非依赖的方式抑制Smad4的转录活性
- Author:
Limin ZHANG
;
Wei LI
;
Xiaoming YANG
- Publication Type:Journal Article
- Keywords:
NLK;
protein kinase;
Smad4;
transcription activity
- From:
Military Medical Sciences
2014;(3):189-192,202
- CountryChina
- Language:Chinese
-
Abstract:
Objective To confirm the interaction betweem NLK and Smad 4 and to explore the effect of NLK on the function of Smad4.Methods Co-immunoprecipitation and GST Pull-down were used to detect the interaction between NLK and Smad4.GST Pull-down was used to map the domain through which Smad 4 interacts with NLK.Luciferase reporter gene assay was used to study the effect of NLK and NLK (KM), the NLK mutant lacking kinase activity , on the transcrip-tion activity of Smad4.In vivo phosphorylation assay was used to detect whether NLK phosphorylated Smad 4 or not.Results The data of Co-immunoprecipitation and GST Pull-down showed that NLK interacted with Smad 4 in vivo and in vitro.The result of GST Pull-down showed that Smad4 interacted with NLK via MH2 domain.The results of luciferase reporter gene assay indicated that both NLK and NLK (KM) inhibited the transcription activity of Smad4.The result of in vivo phospho-rylation assay showed that NLK could not phosphorylate Smad 4 in vivo.Conclusion NLK interacts with Smad4 and inhibits the transcription activity of Smad 4 independent of the kinase activity of NLK .The mechanism through which NLK negatively regulates the transcription of Smad 4 requires further research .
