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Identification and characterization of a soluble cadherin-7 isoform produced by alternative splicing
Rie Kawano1, Noritaka Matsuo, Hideaki Tanaka
1Department of Anatomy, Biology, and Medicine and the Department of Infectious Diseases, Oita Medical University, Hasama-machi, Oita 879-5593, Japan. riekawa@oita-med.ac.jp
Abstract:
We identified an alternative mRNA encoding a novel cadherin-7 isoform by reverse transcriptase-PCR of RNA from day 12 chicken embryos. The alternative mRNA contains 49 bases of insertion in the premembrane region, leading to the substitution of 14 amino acids and the introduction of a premature stop codon. Identification of a 49-bp insertion sequence in the genomic DNA corresponding to the intron of the cadherin-7 gene suggests that alternative splicing is the cause of the alternative mRNA. Transient expression of the variant form in COS-7 or 293 cells produced a soluble protein. Aggregation assays and immunoprecipitation showed that the variant protein interacts with full-length cadherin-7 in vitro and in vivo and inhibits full-length cadherin-7-mediated cell adhesion. Immunohistochemistry revealed that the variant form was strongly expressed in dermomyotomes rather than in migrating neural crest cells, in contrast to the full-length cadherin-7, suggesting differential regulation of splicing and possible roles of variant cadherin-7 in the development of dermomyotomes and other tissues.
Insights
Researchers discovered a new cadherin-7 protein variant in chicken embryos. This variant, produced by alternative splicing, inhibits cell adhesion and is expressed in dermomyotomes, suggesting a role in tissue development.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Cadherin-7 is crucial for cell adhesion during embryonic development.
- Alternative splicing generates protein diversity, impacting cellular functions.
Purpose of the Study:
- To identify and characterize a novel cadherin-7 isoform.
- To investigate the functional consequences of this variant.
Main Methods:
- Reverse transcriptase-PCR to detect alternative mRNA.
- Transient cell expression (COS-7, 293 cells).
- Aggregation assays, immunoprecipitation, and immunohistochemistry.
Main Results:
- Identified an alternative cadherin-7 mRNA with a 49-bp insertion, leading to a premature stop codon.
- The variant protein is soluble, interacts with full-length cadherin-7, and inhibits cell adhesion.
- Variant cadherin-7 is expressed in dermomyotomes, unlike full-length cadherin-7.
Conclusions:
- Alternative splicing produces a soluble, inhibitory cadherin-7 variant.
- Differential expression suggests distinct roles in dermomyotome development.
- The variant may play a specific role in tissue development through regulated splicing.