title: A Thr359Met mutation in factor VII of a patient with a hereditary deficiency causes defective secretion of the molecule.
abstract: We elucidated the genetic basis responsible for factor VII deficiency in an Italian woman with a severe bleeding diathesis. In the allele inherited from the patient's father, we identified a G to A mutation at nucleotide 6070 at the 5' splice site of intron 4 and a G to A substitution at nucleotide 10976 resulting in the Arg353Gln polymorphism. The maternal allele demonstrated a C to T substitution at nucleotide 10994 resulting in Thr359Met. The mutation at nucleotide 6070 alters an invariant GT dinucleotide and disrupts normal mRNA processing. To investigate the mechanism by which Thr359Met reduces factor VIl levels, we expressed wild type factor VII cDNA (FVIIwt) and a mutant factor VII cDNA containing the base substitution resulting in Met359 (FVII359M) in Chinese hamster ovary cells (CHO). In cells transfected with the mutant factor VII cDNA, FVII359M accumulated intracellularly, and no factor VII was detected in the media after 3 hours of chase. The carbohydrate side chains associated with FVII359M were sensitive to Endo H digestion, which indicates that the protein is retained in the endoplasmic reticulum. Analysis of cell lysates also showed that FVII359M was associated with the 78 kD protein corresponding to GRP78/BiP. We conclude that a Thr359Met mutation in factor VII results in a severe secretion defect that probably results from abnormal folding of the molecule.
authors.affiliation: Hematology-Oncology Section, Department of Medicine, Brockton-West Roxbury Department of Veterans Affairs Medical Center.
authors.forename: A A
authors.initials: AA
authors.lastname: Arbini
authors.forename: M
authors.initials: M
authors.lastname: Mannucci
authors.forename: K A
authors.initials: KA
authors.lastname: Bauer
mesh_terms: D000328:Adult; D000483:Alleles; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D005167:Factor VII / Q000737:chemistry / Q000378:metabolism*; D005168:Factor VII Deficiency / Q000235:genetics*; D005260:Female; D006031:Glycosylation; D006360:Heat-Shock Proteins; D006801:Humans; D018832:Molecular Chaperones / Q000378:metabolism; D008969:Molecular Sequence Data; D010375:Pedigree; D017354:Point Mutation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D012326:RNA Splicing
chemical_list: D002352:Carrier Proteins; D003062:Codon; D018076:DNA, Complementary; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D005167:Factor VII