A Thr359met mutation in factor VII of a patient with a hereditary deficiency causes defective secretion of the molecule

Arnaldo A. Arbini, Pier Mannuccio Mannucci, Kenneth A. Bauer

Research output: Contribution to journalArticle

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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 Thr369Met. The mutation at nucleotide 6070 alters an invariant GT dinucleotide and disrupts normal mRNA processing, To investigate the mechanism by which Thr359Met reduces factor VII 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 Th359Met mutation in factor VII results in a severe secretion defect that probably results from abnormal folding of the molecule.

Original languageEnglish
Pages (from-to)5085-5094
Number of pages10
JournalBlood
Volume87
Issue number12
Publication statusPublished - Jun 15 1996

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Factor VII
Mutation
Molecules
Nucleotides
Complementary DNA
Substitution reactions
Factor VII Deficiency
Alleles
RNA Splice Sites
Disease Susceptibility
Cricetulus
Fathers
Endoplasmic Reticulum
Introns
Polymorphism
Digestion
Ovary
Proteins
Mothers
Carbohydrates

ASJC Scopus subject areas

  • Hematology

Cite this

A Thr359met mutation in factor VII of a patient with a hereditary deficiency causes defective secretion of the molecule. / Arbini, Arnaldo A.; Mannucci, Pier Mannuccio; Bauer, Kenneth A.

In: Blood, Vol. 87, No. 12, 15.06.1996, p. 5085-5094.

Research output: Contribution to journalArticle

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