The GCC repeat length in the 5′UTR of MRP1 gene is polymorphic: A functional characterization of its relevance for cystic fibrosis

Abstract

Background: Among the members of the ATP binding cassette transporter superfamily, MRPs share the closest homology with the CFTR protein, which is defective in CF disease. MRP1 has been proposed as a potential modifier gene and/or as novel target for pharmacotherapy of CF to explain the clinical benefits observed in some CF patients treated with the macrolide AZM. The 5′UTR of the MRP 1 gene contains a GCC triplet repeat that could represent a polymorphic site and affect the activity of the promoter. Methods: The MRP1 5′ flanking region was amplified by PCR from 36 CF patients and 100 non-CF subjects and the number of GCC triplets of each allele was determined by sequence and electrophoretic analysis. We performed gene reporter studies in CF airway epithelial cells 16HBE14o-AS3, in basal conditions and in the presence of AZM. Results: We found that the GCC repeat is polymorphic, ranging from 7 to 14 triplets either in CF or in non-CF subjects. Our data are preliminary and have to be confirmed on a larger population of CF subjects. The transcriptional activity of the proximal MRP1 5′ regulatory region revealed no statistically significant correlations between the number of repeats and treatment with AZM. Conclusion: We identified a novel polymorphism in the 5′UTR of MRP 1 gene that provides multiple alleles in a gene relevant for multidrug resistance as well as for CF, determining that this region is transcriptionally active and that this activity does not appear to be influenced by AZM treatment. © 2006 Nicolis et al; licensee BioMed Central Ltd.