Characterization of Allelic and Nucleotide Variation Between the RAGE Gene on Chromosome 6 and a Homologous Pseudogene Sequence to Its 5′ Regulatory Region on Chromosome 3
Implications for Polymorphic Studies in Diabetes
- Academic Unit of Molecular Vascular Medicine, University of Leeds, Leeds General Infirmary, Leeds, U.K.
Abstract
Activation of the receptor for advanced glycation end products (RAGE) appears to be a key mechanism in the pathogenesis of diabetic vascular disease, making RAGE a candidate gene for investigation. RAGE is located in the major histocompatibility complex locus on chromosome 6, which contains a multitude of overlapping and duplicated genes involved predominantly in inflammatory and immune responses. The RAGE 5′ flanking region from −505 in a 5′ direction overlaps with PBX2, a gene that has a pseudogene copy on chromosome 3, making any studies of polymorphisms in this duplicated region potentially fraught with error. In this study we have addressed these issues by confirming RAGE as a predominantly single-copy gene and PBX2 to have two gene copies in the haploid human genome. We have characterized the gene:pseudogene differences between RAGE/PBX2 on chromosome 6 and ΨPBX2 on chromosome 3, which include a change from C to A at position −1139 RAGE/+2298 PBX2, previously reported as a polymorphism. Single chromosome–specific DNA amplification of the duplicated region has clarified five polymorphisms to be on chromosome 3 and one (at −1202 RAGE/+2234 PBX2) to be on chromosome 6. In conclusion, this study provides essential data for the study of RAGE and its genetics.
Footnotes
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Address correspondence and reprint requests to Dr Barry I. Hudson, Academic Unit of Molecular Vascular Medicine, Research School of Medicine, G Floor, Martin Wing, Leeds General Infirmary, Leeds, LS1 3EX, U.K. E-mail: b.hudson{at}leeds.ac.uk.
Received for publication 1 August 2001 and accepted in revised form 1 October 2001. Posted on the World Wide Web at http://www.diabetes.org/diabetes_rapids/ on 9 November 2001.
AGE, advanced glycation end product; DHPLC, denaturing high-performance liquid chromatography; FXIII, Factor XIII; MHC, major histocompatibility complex; PCR, polymerase chain reaction; RAGE, receptor for AGEs; RFLP, restriction fragment–length polymorphism; SSCP, single-strand conformation polymorphism; UTR, untranslated region.
