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A novel c.-22T>C mutation in GALK1 promoter is associated with elevated galactokinase phenotype.
BMC Med Genet. 2009 Mar 24; 10:29.BM

Abstract

BACKGROUND

Many genetic variations of GALK1 have been identified in the patients with galactokinase (GALK1) deficiency. However, the molecular characteristics of GALK1 in individuals with elevated GALK1 activity are relatively unknown.

METHODS

We investigated the relationship between elevated GALK1 activity and the molecular GALK1 gene variations, and the molecular mechanism underlying elevated GALK1 activity. PCR products from 63 subjects, without any attenuation of galactose degradation enzymes, were sequenced to screen for nucleotide alterations in the GALK1 promoter.

RESULTS

Three nucleotide substitutions were identified: c.-179A>G, c.-27A>C, and c.-22T>C. With respect to the c.-22T>C mutation, GALK1 activity in 13 subjects with the T/C or C/C genotype was significantly higher than those in 50 subjects with the T/T genotype (p < 0.001). The dual luciferase reporter assay in Hep3B cells showed that the luciferase activity with the GALK1 promoter with the c.-22C mutant allele increased approximately 2.5-fold, compared to that with the c.-22T. A specific DNA-protein complex was observed in an electrophoretic mobility shift assay, with slightly higher affinity to c.-22C than to c.-22T.

CONCLUSION

The c.-22T>C mutation, which was observed frequently in individuals with elevated GALK1 activity, increased the expression of a reporter gene through enhanced binding of a currently unidentified nuclear protein. These results suggest that the elevated GALK1 activity resulted from enhanced gene expression, due to nucleotide variation within GALK1 promoter.

Authors+Show Affiliations

Ilsong Institute of Life Science, Hallym University, Anyang, Korea. nayadoo@hanmail.netNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19309526

Citation

Park, Hyung-Doo, et al. "A Novel c.-22T>C Mutation in GALK1 Promoter Is Associated With Elevated Galactokinase Phenotype." BMC Medical Genetics, vol. 10, 2009, p. 29.
Park HD, Kim YK, Park KU, et al. A novel c.-22T>C mutation in GALK1 promoter is associated with elevated galactokinase phenotype. BMC Med Genet. 2009;10:29.
Park, H. D., Kim, Y. K., Park, K. U., Kim, J. Q., Song, Y. H., & Song, J. (2009). A novel c.-22T>C mutation in GALK1 promoter is associated with elevated galactokinase phenotype. BMC Medical Genetics, 10, 29. https://doi.org/10.1186/1471-2350-10-29
Park HD, et al. A Novel c.-22T>C Mutation in GALK1 Promoter Is Associated With Elevated Galactokinase Phenotype. BMC Med Genet. 2009 Mar 24;10:29. PubMed PMID: 19309526.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel c.-22T>C mutation in GALK1 promoter is associated with elevated galactokinase phenotype. AU - Park,Hyung-Doo, AU - Kim,Yoon-Kyoung, AU - Park,Kyoung Un, AU - Kim,Jin Q, AU - Song,Young-Han, AU - Song,Junghan, Y1 - 2009/03/24/ PY - 2008/10/07/received PY - 2009/03/24/accepted PY - 2009/3/25/entrez PY - 2009/3/25/pubmed PY - 2009/4/18/medline SP - 29 EP - 29 JF - BMC medical genetics JO - BMC Med. Genet. VL - 10 N2 - BACKGROUND: Many genetic variations of GALK1 have been identified in the patients with galactokinase (GALK1) deficiency. However, the molecular characteristics of GALK1 in individuals with elevated GALK1 activity are relatively unknown. METHODS: We investigated the relationship between elevated GALK1 activity and the molecular GALK1 gene variations, and the molecular mechanism underlying elevated GALK1 activity. PCR products from 63 subjects, without any attenuation of galactose degradation enzymes, were sequenced to screen for nucleotide alterations in the GALK1 promoter. RESULTS: Three nucleotide substitutions were identified: c.-179A>G, c.-27A>C, and c.-22T>C. With respect to the c.-22T>C mutation, GALK1 activity in 13 subjects with the T/C or C/C genotype was significantly higher than those in 50 subjects with the T/T genotype (p < 0.001). The dual luciferase reporter assay in Hep3B cells showed that the luciferase activity with the GALK1 promoter with the c.-22C mutant allele increased approximately 2.5-fold, compared to that with the c.-22T. A specific DNA-protein complex was observed in an electrophoretic mobility shift assay, with slightly higher affinity to c.-22C than to c.-22T. CONCLUSION: The c.-22T>C mutation, which was observed frequently in individuals with elevated GALK1 activity, increased the expression of a reporter gene through enhanced binding of a currently unidentified nuclear protein. These results suggest that the elevated GALK1 activity resulted from enhanced gene expression, due to nucleotide variation within GALK1 promoter. SN - 1471-2350 UR - https://www.unboundmedicine.com/medline/citation/19309526/A_novel_c__22T>C_mutation_in_GALK1_promoter_is_associated_with_elevated_galactokinase_phenotype_ L2 - https://bmcmedgenet.biomedcentral.com/articles/10.1186/1471-2350-10-29 DB - PRIME DP - Unbound Medicine ER -