Mutational spectrum and identification of five novel mutations in G6PC1 gene from a cohort of Glycogen Storage Disease Type 1a

Gene. 2019 Jun 5:700:7-16. doi: 10.1016/j.gene.2019.03.029. Epub 2019 Mar 16.

Abstract

Background: Glycogen storage disease type-1a is an inherited, autosomal recessive disorder caused by mutations in G6PC1 gene leading to deficiency of glucose-6-phosphatase-α specifically in the liver/kidney/intestine.

Patients and methods: DNA of six unrelated Indian GSD-1a patients were screened for mutations in the entire coding region of G6PC1 gene followed by direct DNA sequencing and functional was tested using glucose-6-phosphatase assay.

Results: Mutational screening of GSD-1a patients identified five novel mutations, viz., 1) p.V99Cfs*3, 2) p.G125R, 3) IVS1-2A > T, 4) IVS3 + 39G > A and 5) IVS3 + 42G > A along with three previously reported mutations p.G118D, p.R149Q and p.A331V. Interestingly, each of the p.V99Cfs*3, IVS1-2A > T and p.G118D mutations are identified in two unrelated GSD-1a cases. Further allelic distribution of p.V99Cfs*3 and p.A331V mutations were confirmed by RFLP analysis, consistent with autosomal recessive inheritance. Functional characterization revealed that glucose-6-phosphatase activity was completely abrogated with the mutant proteins p.G125R, p.R149Q, p.G118D, p.A331V and p.V99Cfs*3 than wild-type. However, no significant changes were observed in the expression of mutant constructs at transcription and translation level.

Conclusion: Five novel mutations, p.V99Cfs*3, p.G125R, IVS1-2A > T, IVS3 + 39G > A and IVS3 + 42G > A are reported first time to cause GSD-1a among Indian ethnicity and are not yet reported elsewhere, suggesting separate ethnic founder effects for some mutations among Indian ethnicity.

Keywords: G6PC1; GSD-1a; Glucose-6-phosphatase; Indian ethnicity; Phosphohydrolase assay; Splicing mutation.

MeSH terms

  • Amplified Fragment Length Polymorphism Analysis
  • Child
  • Child, Preschool
  • DNA Mutational Analysis
  • Down-Regulation*
  • Female
  • Gene Frequency
  • Glucose-6-Phosphatase / chemistry
  • Glucose-6-Phosphatase / genetics*
  • Glucose-6-Phosphatase / metabolism*
  • Glycogen Storage Disease Type I / genetics*
  • Glycogen Storage Disease Type I / metabolism
  • HEK293 Cells
  • Humans
  • India
  • Infant
  • Male
  • Models, Molecular
  • Mutation*
  • Protein Conformation
  • Sequence Analysis, DNA / methods*

Substances

  • G6PC1 protein, human
  • Glucose-6-Phosphatase

Supplementary concepts

  • Hepatorenal form of glycogen storage disease