Human Molecular Genetics Advance Access originally published online on February 18, 2009
Human Molecular Genetics 2009 18(9):1624-1632; doi:10.1093/hmg/ddp080
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Origins, distribution and expression of the Duarte-2 (D2) allele of galactose-1-phosphate uridylyltransferase
1 SURE Undergraduate Research Program and 2 Graduate Program in Biochemistry, Cell, and Developmental Biology, Emory University, Atlanta, USA 3 Department of Human Genetics, Emory University School of Medicine, Atlanta, GA 30322, USA and 4 Division of Medical Genetics, Department of Human Genetics, Emory University School of Medicine, Decatur, GA 30033, USA
* To whom correspondence should be addressed at: Department of Human Genetics, Rm. 325.2 Whitehead Bldg., 615 Michael St., Atlanta, GA 30322, USA. Tel: +1 4047273924; Fax: +1 4047273949; Email: jfridov{at}emory.edu
Received January 8, 2009; Accepted February 16, 2009
Duarte galactosemia is a mild to asymptomatic condition that results from partial impairment of galactose-1-phosphate uridylyltransferase (GALT). Patients with Duarte galactosemia demonstrate reduced GALT activity and carry one profoundly impaired GALT allele (G) along with a second, partially impaired GALT allele (Duarte-2, D2). Molecular studies reveal at least five sequence changes on D2 alleles: a p.N314D missense substitution, three intronic base changes and a 4 bp deletion in the 5' proximal sequence. The four non-coding sequence changes are unique to D2. The p.N314D substitution, however, is not; it is found together with a silent polymorphism, p.L218(TTA), on functionally normal Duarte-1 alleles (D1, also called Los Angeles or LA alleles). The HapMap database reveals that p.N314D is a common human variant, and cross-species comparisons implicate D314 as the ancestral allele. The p.N314D substitution is also functionally neutral in mammalian cell and yeast expression studies. In contrast, the 4 bp 5' deletion characteristic of D2 alleles appears to be functionally impaired in reporter gene transfection studies. Here we present allele-specific qRT–PCR evidence that D2 alleles express less mRNA in vivo than their wild-type counterparts; the difference is small but statistically significant. Furthermore, we characterize the prevalence of the 4 bp deletion in GG, NN and DG populations; the deletion appears exclusive to D2 alleles. Combined, these data strongly implicate the 4 bp 5' deletion as a causal mutation in Duarte galactosemia and suggest that direct tests for this deletion, as proposed here, could enhance or supplant current tests, which define D2 alleles on the basis of the presence and absence of linked coding sequence polymorphisms.
These authors contributed equally to this work.