Synonymous nucleotide polymorphisms influence Dombrock blood group protein expression in K562 cells. (11th October 2013)

• Record Type: Journal Article
• Authors: de, Cécile Durousseau; Silvy, Monique; Halverson, Gregory; Chiaroni, Jacques; Bailly, Pascal; Chapel‐Fernandes, Sylvie
• Is Part Of: British journal of haematology. Volume 164:Number 1(2014:Jan.)
• Page Start: 131
• Page End: 141
• Publication Date: 2013-10-11
• Subjects: Hematology -- Periodicals; Blood -- Diseases -- Periodicals; 616.15
• DOI: 10.1111/bjh.12597
• Languages: English
• ISSNs: 0007-1048

Abstract

<abstract abstract-type="main" id="bjh12597-abs-0001"> <title>Summary</title> <p>To gain further insight into <italic>ART</italic><italic>4</italic> (<italic>DO</italic>) gene alleles (<italic>DO</italic><italic>*</italic><italic>A</italic>, <italic>DO</italic><italic>*</italic><italic>JO</italic><italic>1, </italic><italic>DO</italic><italic>*</italic><italic>A‐WL</italic><italic>, </italic><italic>DO</italic><italic>*</italic><italic>DOYA</italic><italic>, </italic><italic>DO</italic><italic>*</italic><italic>B</italic><italic>, </italic><italic>DO</italic><italic>*</italic><italic>B‐WL</italic><italic>, </italic><italic>DO</italic><italic>*</italic><italic>B‐SH</italic><italic>‐Q149K, DO*B‐(WL)‐I175N, DO*HY1, DO*HY2, DO*DOMR)</italic> and evaluate the impact of synonymous nucleotide polymorphisms on protein expression and mRNA accumulation of <italic>DO*A‐HA</italic>, <italic> DO*A‐SH</italic> and <italic>DO*B‐SH</italic> alleles, human erythroleukaemic K562 cells were transducted with variant <italic>DO</italic>‐lentiviral particles and analysed by flow cytometry and quantitative reverse transcription polymerase chain reaction. Monoclonal antibody (MoAb) detection of <italic>DO*A‐HA</italic> and <italic>DO*JO1</italic> transductants was lower than <italic>DO*A</italic> transductants, while detection of <italic>DO*A‐SH</italic>, <italic> DO*A‐WL</italic> and <italic>DO*DOYA</italic> transductants was higher. Variant <italic>DO*B</italic> alleles, i.e. <italic>DO*B‐SH</italic>, <italic> DO*B‐WL</italic>, <italic> DO*HY1</italic>, <italic> DO*HY2</italic> and <italic>DO*DOMR</italic>, showed reduced MoAb binding. The unexpected modifications of protein expression of the <italic>DO*A‐HA, DO*A‐SH</italic> and <italic>DO*B‐SH</italic> alleles that differ from the <italic>DO*A</italic> or <italic>DO*B</italic> alleles by a single synonymous polymorphism were abolished by reversion, thus implying involvement of these polymorphisms. Depending on the Leu208 codon used, detection level ranged from 1 to 4·14. In the variant alleles resulting from single synonymous polymorphism, mRNA accumulation correlated roughly with MoAbs detection levels, suggesting post‐transcriptional regulation. Other than a few reports involving aberrant splicing, the experiments described herein provide the first evidence that synonymous nucleotide polymorphisms can influence Dombrock blood group expression. Such polymorphisms should be taken into account for molecular screening and potential impact on transfusion.</p> </abstract>