title: Type I Glanzmann thrombasthenia caused by an apparently silent beta3 mutation that results in aberrant splicing and reduced beta3 mRNA.
abstract: We report a novel genetic defect in a patient with type I Glanzmann thrombasthenia. Flow cytometry analysis revealed undetectable levels of platelet glycoproteins alphaIIb and beta3, although residual amounts of both proteins were detectable in immunoblotting analysis. Sequence analysis of reversely transcribed platelet beta3 mRNA showed a 100-base pair deletion in the 3'-boundary of exon 11, that results in a frame shift and appearance of a premature STOP codon. Analysis of the corresponding genomic DNA fragment revealed the presence of a homozygous C1815T transition in exon 11. The mutation does not change the amino acid residue but it creates an ectopic consensus splice donor site that is used preferentially, causing splicing out of part of exon 11. The parents of the proband, heterozygous for this mutation, were asymptomatic and had reduced platelet content of alphaIIbbeta3. PCR-based relative quantification of beta3 mRNA failed to detect the mutant transcript in the parents and showed a marked reduction in the patient. The results suggest that the thrombasthenic phenotype is, mainly, the result of the reduced availability of beta3-mRNA, most probably due to activation of the nonsense-mediated mRNA decay mechanism. They also show the convenience of analyzing both genomic DNA and mRNA, in order to ascertain the functional consequences of single nucleotide substitutions.
authors.affiliation: Department of Pathophysiology and Human Molecular Genetics Centro de Investigaciones Biológicas (CSIC) Ramiro de Maeztu, Madrid, Spain.
authors.forename: Jingli
authors.initials: J
authors.lastname: Xie
authors.forename: Dina
authors.initials: D
authors.lastname: Pabón
authors.forename: Asier
authors.initials: A
authors.lastname: Jayo
authors.forename: Nora
authors.initials: N
authors.lastname: Butta
authors.forename: Consuelo
authors.initials: C
authors.lastname: González-Manchón
mesh_terms: D000483:Alleles; D017398:Alternative Splicing; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001792:Blood Platelets / Q000378:metabolism; D016466:CHO Cells; D002675:Child, Preschool; D018389:Codon, Nonsense; D018388:Codon, Terminator; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005091:Exons; D005192:Family Health; D005220:Fathers; D005260:Female; D005340:Fibrinogen / Q000737:chemistry; D005434:Flow Cytometry; D016368:Frameshift Mutation; D005821:Genetic Techniques; D006720:Homozygote; D006801:Humans; D047468:Immunoprecipitation; D039661:Integrin beta3 / Q000235:genetics*; D008297:Male; D051379:Mice; D008957:Models, Genetic; D009035:Mothers; D009154:Mutation; D010641:Phenotype; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013915:Thrombasthenia / Q000209:etiology* / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; D018389:Codon, Nonsense; D018388:Codon, Terminator; D039661:Integrin beta3; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D012333:RNA, Messenger; D005340:Fibrinogen; D004247:DNA