title: Quality control of fibrinogen secretion in the molecular pathogenesis of congenital afibrinogenemia.
abstract: Congenital afibrinogenemia is a rare bleeding disorder characterized by the absence in circulation of fibrinogen, a hexamer composed of two sets of three polypeptides (Aalpha, Bbeta and gamma). Each polypeptide is encoded by a distinct gene, FGA, FGB and FGG, all three clustered in a region of 50 kb on 4q31. A subset of afibrinogenemia mutations has been shown to specifically impair fibrinogen secretion, but the underlying molecular mechanisms remained to be elucidated. Here, we show that truncation of the seven most C-terminal residues (R455-Q461) of the Bbeta chain specifically inhibits fibrinogen secretion. Expression of additional mutants and structural modelling suggests that neither the last six residues nor R455 is crucial per se for secretion, but prevent protein misfolding by protecting hydrophobic residues in the betaC core. Immunofluorescence and immuno-electron microscopy studies indicate that secretion-impaired mutants are retained in a pre-Golgi compartment. In addition, expression of Bbeta, gamma and angiopoietin-2 chimeric molecules demonstrated that the betaC domain prevents the secretion of single chains and complexes, whereas the gammaC domain allows their secretion. Our data provide new insight into the mechanisms accounting for the quality control of fibrinogen secretion and confirm that mutant fibrinogen retention is one of the pathological mechanisms responsible for congenital afibrinogenemia.
authors.affiliation: Department of Genetic Medicine and Development, Swiss Institute of Bioinformatics, University Medical Centre, Geneva, Switzerland.
authors.forename: Dung
authors.initials: D
authors.lastname: Vu
authors.forename: Corinne
authors.initials: C
authors.lastname: Di Sanza
authors.forename: Dorothée
authors.initials: D
authors.lastname: Caille
authors.forename: Philippe
authors.initials: P
authors.lastname: de Moerloose
authors.forename: Holger
authors.initials: H
authors.lastname: Scheib
authors.forename: Paolo
authors.initials: P
authors.lastname: Meda
authors.forename: Marguerite
authors.initials: M
authors.lastname: Neerman-Arbez
mesh_terms: D000347:Afibrinogenemia / Q000151:congenital / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D017931:DNA Primers; D005340:Fibrinogen / Q000235:genetics* / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D016253:Microscopy, Immunoelectron; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D017510:Protein Folding
chemical_list: C115499:BBeta fibrinogen; D017931:DNA Primers; D005340:Fibrinogen