The localization of FGFR3 mutations causing thanatophoric dysplasia type I differentially affects phosphorylation, processing and ubiquitylation of the receptor.

Abstract

Recurrent missense fibroblast growth factor receptor 3 (FGFR3) mutations have been ascribed to skeletal dysplasias of variable severity including the lethal neonatal thanatophoric dysplasia types I (TDI) and II (TDII). To elucidate the role of activating mutations causing TDI on receptor trafficking and endocytosis, a series of four mutants located in different domains of the receptor were generated and transiently expressed. The putatively elongated X807R receptor was identified as three isoforms. The fully glycosylated mature isoform was constitutively but mildly phosphorylated. Similarly, mutations affecting the extracellular domain (R248C and Y373C) induced moderate constitutive receptor phosphorylation. By contrast, the K650M mutation affecting the tyrosine kinase 2 (TK2) domain produced heavy phosphorylation of the nonglycosylated and mannose-rich isoforms that impaired receptor trafficking through the Golgi network. This resulted in defective expression of the mature isoform at the cell surface. Normal processing was rescued by tyrosine kinase inhibitor treatment. Internalization of the R248C and Y373C mutant receptors, which form stable disulfide-bonded dimers at the cell surface was less efficient than the wild-type, whereas ubiquitylation was markedly increased but apparently independent of the E3 ubiquitin-ligase casitas B-lineage lymphoma (c-Cbl). Constitutive phosphorylation of c-Cbl by the K650M mutant appeared to be related to the intracellular retention of the receptor. Therefore, although mutation K650M affecting the TK2 domain induces defective targeting of the overphosphorylated receptor, a different mechanism characterized by receptor retention at the plasma membrane, excessive ubiquitylation and reduced degradation results from mutations that affect the extracellular domain and the stop codon.

• Authors: Jacky Bonaventure, Linda Gibbs, William C Horne, Roland Baron
• Journal: The FEBS journal (FEBS J) Vol. 274 Issue 12 Pg. 3078-93 (Jun 2007) ISSN: 1742-464X [Print] England
• PMID: 17509076 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Codon, Terminator
• Protein Isoforms
• Brefeldin A
• Proto-Oncogene Proteins c-cbl
• FGFR3 protein, human
• Receptor, Fibroblast Growth Factor, Type 3
• TYK2 Kinase
• TYK2 protein, human
• Mannose

Topics

• Brefeldin A (pharmacology)
• Cell Line
• Cell Membrane (metabolism)
• Codon, Terminator
• Endocytosis
• Glycosylation
• Golgi Apparatus (metabolism)
• Humans
• Mannose (metabolism)
• Mutagenesis, Site-Directed
• Mutation, Missense
• Phosphorylation
• Protein Isoforms (genetics, metabolism)
• Protein Structure, Tertiary
• Protein Transport
• Proto-Oncogene Proteins c-cbl (metabolism)
• Receptor, Fibroblast Growth Factor, Type 3 (genetics, metabolism)
• TYK2 Kinase (metabolism)
• Thanatophoric Dysplasia (genetics)