Identification of 11 novel mutations in 49 Korean patients with mucopolysaccharidosis type II.

Mucopolysaccharidosis type II (MPS II) or Hunter syndrome is a rare lysosomal storage disorder caused by a deficiency of iduronate-2-sulfatase (IDS). As MPS II is X-linked, patients are usually males with heterogeneous mutations ranging from point mutations to gross deletions and recombination. In 2003, we reported a mutation analysis of 25 patients with MPS II. In this study, 31 mutations in another 49 Korean patients (45 families) with MPS II are reported: 12 missense, nine deletions, four splicing, two nonsense, two insertions, one deletion/insertion, and IDS-IDS2 recombination mutations. Among these mutations, 11 were novel ones (4 missense mutations: Ser61Pro, Pro97Arg, Pro228Ala, and Pro261Ala; 5 deletions: c.344delA, c.420delG, c.768delT, c.1112delC and c.1402delC; 1 deletion/insertion: c.1222delinsTA; and 1 insertion mutation: c.359_360insATCC). The IDS-IDS2 recombination mutations were most frequently observed; all patients with this mutation had the severe MPS II phenotype. However, most of the patients (5/7) with the G374G splicing mutation had an attenuated phenotype, except for two sibling cases with the severe phenotype. Except for a few recurrent mutations such as the G374G, R443X, L522P, and recombination mutations, each patient had a unique individual mutation. Therefore, careful interpretation of genotype-phenotype correlations is warranted.

Cloning and sequence analysis of Gpdh in Callosobruchus chinensis (Coleoptera: Bruchidae).

The Sn-Glycerol-3-phosphate dehydrogenase (GPDH: NAD+ 2-oxidoreductase, EC 1.1.1.8) gene of C. chinensis was cloned and its nucleotide sequence was analyzed. The gene was obtained by screening a genomic library with Drosophila melanogaster Gpdh and PCR amplification. The 5,126 bp gene obtained is comprised of one 5' untranslated region, eight exons, seven introns, and three 3' untranslated regions. Comparison of Gpdh of D. melanogaster with that of C. chinensis showed a 89.9% identity in the coding region, 70% in the intron, 79% in the entire nucleotide sequence, and 83.2% in the deduced amino acid sequence. The transcription initiation site is located 33 nucleotides upstream of the initiation codon, and the sequence analysis of the promoter region showed TATA and CAAT boxes at the 5' end. The stop codon (TAA) and polyadenylation signal (AATAAA) are located at the 3' end of each of the exons 6 to 8. These findings show that GPDH isozymes in C. chinensis are produced by the alternative processing of 3' exons. The occurrence of the three transcripts was proven by RT-PCR using synthetic oligonucleotides complementary to the predicted unique 3' regions. Compared to the D. melanogaster GPDH isozymes, GPDH-1, -2, and -3, C. chinensis GPDH showed 83.6%, 83%, and 84% identities, respectively.