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Published on: December 29, 2009
[Screening of proteins interacting with Dishevelled2 in mouse 11.5dpc embryo library]
Ser-Sue Ng1, Yong-Gong Zhai, Liang Han
1Tsinghua Institute of Genome Research, Department of Biological Sciences and Biotechnology, Institute of Biomedicine, Tsinghua University, Beijing 100084, China.
Abstract:
Dishevelled proteins are multifunctional and highly conserved. These proteins are also required for the specification of cell fate and polarity by secreted Wnt proteins. To investigate the molecular mechanism of Dishevelled in mediating Wnt signal transduction, a mouse 11.5dpc embryo library was screened by yeast-two-hybrid system to find mouse Dishevelled2 DEP domain and C-terminal interacting proteins. 15 possitive clones were identified from 4.1 x 10(6) transformants. The DNA sequences of the positive AD/library plasmids were determined. The BLAST results revealed that one of the positive clones contained N-terminus cDNA fragments (amino acids 6-122) of Gli3 protein. The interaction between Dv12 and Gli3 detected by yeast two-hybrid system suggests that Gli3 might play a role in some biological processes with Dishevelled.
Insights
Researchers identified a novel interaction between Dishevelled (Dvl) proteins and Gli3, suggesting Gli3 plays a role in Wnt signaling pathways. This finding advances understanding of cell fate and polarity determination.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- Dishevelled (Dvl) proteins are crucial, conserved mediators of Wnt signaling, essential for cell fate and polarity.
- Understanding the molecular mechanisms of Dvl in Wnt signal transduction is vital for developmental biology.
Purpose of the Study:
- To investigate the molecular mechanisms of Dishevelled in mediating Wnt signal transduction.
- To identify interacting proteins of the mouse Dishevelled2 DEP domain and C-terminus.
Main Methods:
- Screening of a mouse 11.5dpc embryo library using a yeast-two-hybrid system.
- DNA sequencing and BLAST analysis of positive clones to identify interacting proteins.
Main Results:
- 15 positive clones were identified from 4.1 x 10^6 transformants.
- One positive clone contained N-terminus cDNA fragments of the Gli3 protein.
- The yeast-two-hybrid system detected an interaction between Dishevelled 2 (Dv12) and Gli3.
Conclusions:
- Gli3 interacts with Dishevelled 2, suggesting a potential role for Gli3 in Dishevelled-mediated biological processes.
- This interaction may offer new insights into the regulation of Wnt signaling pathways and developmental processes.
