Identification of three novel mutations in the PIG-A gene in paroxysmal nocturnal haemoglobinuria (PNH) patients.
Paroxysmal nocturnal haemoglobinuria (PNH) is an acquired haemolytic disorder caused by the absence of glycosyl phosphatidylinositol (GPI)-anchored surface proteins resulting from a defect in one step of GPI-anchor biosynthesis. Recent analysis has shown that mutations at the PIG-A (phosphatidylinositoglycan-class A) gene are responsible for GPI-anchor deficiency in all PNH patients. In the current study, we describe three new mutations of the PIG-A gene in Italian patients with PNH. The analysis has been performed by RNA/single-strand conformation polymorphism using genomic DNA purified from nucleated peripheral blood cells. An abnormal pattern of migration of polymerase chain reaction amplified fragments containing exons 2 and 5 was observed. Sequencing analysis led to the identification of three mutations: a transversion C-to-A creating a stop codon (Y98X), an A insertion at position 460 (460insA), and a C deletion (1114delC). All the mutations cause a premature termination of the translation of the PIG-A protein.
Savoia
A
A
Servizio di Genetica Medica, I.R.C.C.S. Ospedale Casa Sollievo della Sofferenza, San Giovanni Rotondo, Foggia, Italy.
Ianzano
L
L
Lunardi
C
C
De Sandre
G
G
Carotenuto
M
M
Musto
P
P
Zelante
L
L
D001483:Base Sequence; D003062:Codon; D004247:DNA / Q000097:blood; D017931:DNA Primers; D004912:Erythrocytes / Q000378:metabolism; D005091:Exons; D017261:Glycosylphosphatidylinositols / Q000096:biosynthesis / Q000235:genetics*; D006457:Hemoglobinuria, Paroxysmal / Q000235:genetics*; D006801:Humans; D007558:Italy; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion
D003062:Codon; D017931:DNA Primers; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; C080086:phosphatidylinositol glycan-class A protein; D004247:DNA