+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Mutation screening of the entire coding regions of the TSC1 and the TSC2 gene with the protein truncation test (PTT) identifies frequent splicing defects

Mutation screening of the entire coding regions of the TSC1 and the TSC2 gene with the protein truncation test (PTT) identifies frequent splicing defects

Human Mutation 14(5): 401-411

Mutation analyses in tuberous sclerosis (TSC) have reported a wide variety of disease-causing aberrations in the two known predisposing genes, TSC1 and TSC2 on chromosomes 9q34 and 16p13, comprising mainly small mutations distributed over the entire genes. So far, all known TSC1 mutations as well as the majority of TSC2 mutations truncate the proteins hamartin and tuberin, respectively. We describe for the first time an RNA-based screening of the entire coding regions of both TSC genes for truncating mutations applying the protein truncation test (PTT). Simultaneous investigation of both TSC genes in a group of 48 unassigned TSC patients, which were previously tested to exclude large intragenic TSC2 rearrangements, revealed aberrant migrating polypeptides resulting from truncating mutations in nine TSC1 cases and in 16 TSC2 cases while three TSC2 cases showed enlarged proteins. TSC1 mutations include two nonsense mutations, four insertions, and three splice mutations. Nineteen mutations identified in TSC2 were composed of four different nonsense mutations in five patients, one deletion, one insertion, and seven different splicing aberrations due to at least eight different mutations found in 12 patients. Additional predicted truncating mutations according to PTT without possible identification of the causative alteration allowed assignment to TSC1 in one and TSC2 in seven cases. Twelve patients without abnormalities in the PTT are assumed to harbor missense mutations, probably in TSC2. The high proportion of TSC2 splicing aberrations strengthens the importance of intronic disease-causing mutations and the application of RNA-based screening methods to confirm their consequences.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 011029509

Download citation: RISBibTeXText

PMID: 10533066

DOI: 10.1002/(sici)1098-1004(199911)14:5<401::aid-humu6>3.0.co;2-r

Related references

Molecular diagnosis in tuberous sclerosis Mutation screening of the TSC1 and TSC2 gene by using the protein truncation test. European Journal of Human Genetics 6(Suppl. 1): 147, 1998

Mutations in the TSC2 gene: analysis of the complete coding sequence using the protein truncation test (PTT). Human Molecular Genetics 6(9): 1409-1414, 1997

Tuberous sclerosis type 2 RNA based detection of complex splicing aberrations in the TSC2 gene by the protein truncation test. American Journal of Human Genetics 65(4): A376, 1999

Application of the protein truncation test (PTT) for the detection of tuberculosis sclerosis complex type 1 and 2 (TSC1 and TSC2) mutations. Methods in Molecular Biology 217: 329-344, 2003

Mutation screening of the Tuberous Sclerosis gene 1 by protein truncation test. American Journal of Human Genetics 65(4): A284, 1999

Distribution of Tsc1 protein detected by immunohistochemistry in various normal rat tissues and the renal carcinomas of Eker rat: detection of limited colocalization with Tsc1 and Tsc2 gene products in vivo. Laboratory Investigation; a Journal of Technical Methods and Pathology 80(9): 1347-1359, 2000

Mutation screening of TSC1 and TSC2 genes in Chinese Han children with tuberous sclerosis complex. Genetics and Molecular Research 13(1): 2102-2106, 2014

Tuberous sclerosis locus assignment and TSC2 mutation detection by Southern blot, loss of heterozygosity analysis and Protein Truncation Test. American Journal of Human Genetics 59(4 Suppl. ): A399, 1996

Type VII collagen gene mutation screening in recessive dystrophic epidermolysis bullosa using the protein truncation test. Journal of Dermatological Science 16(Suppl. 1): S146, 1998

TSC1 and TSC2 mutations in tuberous sclerosis, the associated phenotypes and a model to explain observed TSC1/TSC2 frequency ratios. European Journal of Pediatrics 161(7): 393-402, 2002

Analysis of 65 tuberous sclerosis complex (TSC) patients by TSC2 DGGE, TSC1/TSC2 MLPA, and TSC1 long-range PCR sequencing, and report of 28 novel mutations. Human Mutation 26(4): 374-383, 2005

The expression of hamartin, the product of the TSC1 gene, in normal human tissues and in TSC1- and TSC2-linked angiomyolipomas. Modern Pathology 12(5): 539-545, 1999

The TSC1-TSC2 complex consists of multiple TSC1 and TSC2 subunits. Bmc Biochemistry 13: 18, 2012

Comparative mutation detection screening of the type VII collagen gene (COL7A1) using the protein truncation test, fluorescent chemical cleavage of mismatch, and conformation sensitive gel electrophoresis. Journal of Investigative Dermatology 113(4): 673-686, 1999