PUBLICATION
Structure-function analysis of the EGF-CFC family member Cripto identifies residues essential for nodal signalling
- Authors
- Minchiotti, G., Manco, G., Parisi, S., Lago, C.T., Rosa, F., and Persico, M.G.
- ID
- ZDB-PUB-011126-4
- Date
- 2001
- Source
- Development (Cambridge, England) 128(22): 4501-4510 (Journal)
- Registered Authors
- Rosa, Frederic
- Keywords
- Cripto; nodal signalling; zebrafish; one-eyed pinhead; structure-function
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Binding Sites
- Computer Simulation
- Cysteine
- Embryonic Induction*
- Epidermal Growth Factor
- Genetic Complementation Test
- Homeodomain Proteins/genetics
- Homeodomain Proteins/metabolism
- Models, Molecular
- Molecular Sequence Data
- Neoplasm Proteins/administration & dosage
- Neoplasm Proteins/genetics
- Neoplasm Proteins/metabolism*
- Point Mutation
- Protein Binding
- Protein Structure, Tertiary
- Recombinant Proteins/administration & dosage
- Recombinant Proteins/metabolism
- Sequence Homology, Amino Acid
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Zebrafish/embryology*
- Zebrafish Proteins/administration & dosage
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism*
- PubMed
- 11714675 Full text @ Development
Citation
Minchiotti, G., Manco, G., Parisi, S., Lago, C.T., Rosa, F., and Persico, M.G. (2001) Structure-function analysis of the EGF-CFC family member Cripto identifies residues essential for nodal signalling. Development (Cambridge, England). 128(22):4501-4510.
Abstract
cripto is the founding member of the family of EGF-CFC genes, a class of extracellular factors essential for early vertebrate development. In this study we show that injection of Cripto recombinant protein in mid to late zebrafish Maternal-Zygotic one-eyed pinhead (MZoep) blastulae was able to fully rescue the mutant phenotype, thus providing the first direct evidence that Cripto activity can be added extracellularly to recover oep-encoded function in zebrafish early embryos. Moreover, 15 point mutations and two deletion mutants were generated to assess in vivo their functional relevance by comparing the ability of cripto wild-type and mutant RNAs to rescue the zebrafish MZoep mutant. From this study we concluded that the EGF-CFC domain is sufficient for Cripto biological activity and identified ten point mutations with a functional defective phenotype, two of which, located in the EGF-like domain, correspond to loss-of-function mutations. Finally, we have developed a three-dimensional structural model of Cripto protein and used it as a guide to predict amino acid residues potentially implicated in protein-protein interaction.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping