Novel truncating mutations of the CHM gene in Chinese patients with choroideremia

Shea Ping Yip, Tsz Shan Cheung, Man Yu Chu, Suk Chun Cheung, Kam Wah Leung, Kin Ping Tsang, Stephen T.S. Lam, Chi Ho To

Research output: Journal article publicationJournal articleAcademic researchpeer-review

11 Citations (Scopus)

Abstract

Purpose: Choroideremia (CHM) is an X-linked retinal degenerative disorder caused by mutations in the CHM gene. The mutations result in malfunction of the Rab escort protein 1 (REP-1). In this study, mutational analysis of the CHM gene was performed on five Chinese families clinically diagnosed with CHM. Methods: Denaturing high performance liquid chromatography was used for mutation screening for all 15 exons and flanking intron regions of the CHM gene. Mutations were confirmed and characterized with DNA sequencing. Second samples were later collected for extraction of mRNA and proteins from leukocytes. A non-radioactive protein truncation test (PTT) was developed and used to characterize the truncating nature of the mutations. Immunoblot analysis of proteins extracted from leukocytes was also performed. Results: Five mutations were identified in these five families, each with one distinct mutation: three frameshift, one nonsense, and one splicing. Two of these were novel mutations: c.627dupA in exon 5 and c.703- 1G>C in intron 5. The truncating nature of the mutations was experimentally proved by PTT for four families with second samples collected. In particular, c.703- 1G>C spliced exon 5 directly to exon 7 and deleted the entire exon 6 from the transcript. Direct immunoblot analysis failed to detect REP-1 in males affected by CHM, but demonstrated its presence in female carriers and homozygous normal females. Conclusions: This is the first study reporting mutations in the CHM gene in Chinese farnflies. Mutational analysis was performed at the DNA, mRNA and protein levels. Five truncating mutations were found, and two of these were novel.
Original languageEnglish
Pages (from-to)2183-2193
Number of pages11
JournalMolecular Vision
Volume13
Publication statusPublished - 27 Nov 2007

ASJC Scopus subject areas

  • Ophthalmology

Cite this