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Mutation analysis of the cathepsin C gene in Indian families with Papillon-Lefèvre syndrome.

Selvaraju V, Markandaya M, Prasad PV, Sathyan P, Sethuraman G, Srivastava SC, Thakker N, Kumar A - BMC Med. Genet. (2003)

Bottom Line: This study reported three novel nonsense mutations in three Indian families.These novel nonsense mutations are predicted to produce truncated dipeptidyl-peptidase I causing PLS phenotype in these families.A review of the literature along with three novel mutations reported here showed that the total number of mutations in the CTSC gene described to date is 41 with 17 mutations being located in exon 7.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Reproduction Development and Genetics, Indian Institute of Science, Bangalore, India. selvacell@rediffmail.com

ABSTRACT

Background: PLS is a rare autosomal recessive disorder characterized by early onset periodontopathia and palmar plantar keratosis. PLS is caused by mutations in the cathepsin C (CTSC) gene. Dipeptidyl-peptidase I encoded by the CTSC gene removes dipeptides from the amino-terminus of protein substrates and mainly plays an immune and inflammatory role. Several mutations have been reported in this gene in patients from several ethnic groups. We report here mutation analysis of the CTSC gene in three Indian families with PLS.

Methods: Peripheral blood samples were obtained from individuals belonging to three Indian families with PLS for genomic DNA isolation. Exon-specific intronic primers were used to amplify DNA samples from individuals. PCR products were subsequently sequenced to detect mutations. PCR-SCCP and ASOH analyses were used to determine if mutations were present in normal control individuals.

Results: All patients from three families had a classic PLS phenotype, which included palmoplantar keratosis and early-onset severe periodontitis. Sequence analysis of the CTSC gene showed three novel nonsense mutations (viz., p.Q49X, p.Q69X and p.Y304X) in homozygous state in affected individuals from these Indian families.

Conclusions: This study reported three novel nonsense mutations in three Indian families. These novel nonsense mutations are predicted to produce truncated dipeptidyl-peptidase I causing PLS phenotype in these families. A review of the literature along with three novel mutations reported here showed that the total number of mutations in the CTSC gene described to date is 41 with 17 mutations being located in exon 7.

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Three novel nonsense mutations in the CTSC gene. The numbering of the wild type sequence is based upon the cDNA sequence of the CTSC gene (GenBank accession # NM-001814). (A) Pedigree diagram of the family IISC-PLS1. DNA samples from individuals II-4, II-5, III-1 and III-2 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents II-4 and II-5 and homozygous patients III-1 and III-2 are shown on the right. Arrows indicate the145C>T nucleotide change. (B) Pedigree diagram of the family IISC-PLS2. DNA samples from individuals III-3, III-4, IV-1, IV-3, IV-4 and IV-5 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents III-3 and III-4, and homozygous patients IV-3 and IV-4 are shown on the right. Arrows indicate the 205C>T nucleotide change. (C) Pedigree diagram of the family IISC-PLS3. DNA sample from only individual II-2 was analysed from this family. Direct sequencing of the PCR product from the patient II-2 is shown on the right. Arrow indicates the 912 C>A change.
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Figure 2: Three novel nonsense mutations in the CTSC gene. The numbering of the wild type sequence is based upon the cDNA sequence of the CTSC gene (GenBank accession # NM-001814). (A) Pedigree diagram of the family IISC-PLS1. DNA samples from individuals II-4, II-5, III-1 and III-2 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents II-4 and II-5 and homozygous patients III-1 and III-2 are shown on the right. Arrows indicate the145C>T nucleotide change. (B) Pedigree diagram of the family IISC-PLS2. DNA samples from individuals III-3, III-4, IV-1, IV-3, IV-4 and IV-5 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents III-3 and III-4, and homozygous patients IV-3 and IV-4 are shown on the right. Arrows indicate the 205C>T nucleotide change. (C) Pedigree diagram of the family IISC-PLS3. DNA sample from only individual II-2 was analysed from this family. Direct sequencing of the PCR product from the patient II-2 is shown on the right. Arrow indicates the 912 C>A change.

Mentions: We initially screened PCR products of all seven exons of the CTSC gene for mutation(s) in one patient from each family using DNA sequencing technique (Fig. 2). Once the mutation was detected, it was confirmed in other members of the family. Sequence analysis of the exon 1 PCR product from the patient III-1 of the family IISC-PLS1 showed a nucleotide change in a homozygous state at position 145 from C>T (c.145C>T) which introduced a premature stop codon in the exon 1 (p.Q49X) (Fig. 2A). This change was also present in affected sibling III-2 in a homozygous state, whereas both parents were heterozygous for this change (Fig. 2A). PCR-SSCP analysis showed that this change was not present in 60 normal chromosomes (data not shown).


Mutation analysis of the cathepsin C gene in Indian families with Papillon-Lefèvre syndrome.

Selvaraju V, Markandaya M, Prasad PV, Sathyan P, Sethuraman G, Srivastava SC, Thakker N, Kumar A - BMC Med. Genet. (2003)

Three novel nonsense mutations in the CTSC gene. The numbering of the wild type sequence is based upon the cDNA sequence of the CTSC gene (GenBank accession # NM-001814). (A) Pedigree diagram of the family IISC-PLS1. DNA samples from individuals II-4, II-5, III-1 and III-2 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents II-4 and II-5 and homozygous patients III-1 and III-2 are shown on the right. Arrows indicate the145C>T nucleotide change. (B) Pedigree diagram of the family IISC-PLS2. DNA samples from individuals III-3, III-4, IV-1, IV-3, IV-4 and IV-5 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents III-3 and III-4, and homozygous patients IV-3 and IV-4 are shown on the right. Arrows indicate the 205C>T nucleotide change. (C) Pedigree diagram of the family IISC-PLS3. DNA sample from only individual II-2 was analysed from this family. Direct sequencing of the PCR product from the patient II-2 is shown on the right. Arrow indicates the 912 C>A change.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC183830&req=5

Figure 2: Three novel nonsense mutations in the CTSC gene. The numbering of the wild type sequence is based upon the cDNA sequence of the CTSC gene (GenBank accession # NM-001814). (A) Pedigree diagram of the family IISC-PLS1. DNA samples from individuals II-4, II-5, III-1 and III-2 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents II-4 and II-5 and homozygous patients III-1 and III-2 are shown on the right. Arrows indicate the145C>T nucleotide change. (B) Pedigree diagram of the family IISC-PLS2. DNA samples from individuals III-3, III-4, IV-1, IV-3, IV-4 and IV-5 were analysed from this family. Direct sequencing of the PCR products from both heterozygous parents III-3 and III-4, and homozygous patients IV-3 and IV-4 are shown on the right. Arrows indicate the 205C>T nucleotide change. (C) Pedigree diagram of the family IISC-PLS3. DNA sample from only individual II-2 was analysed from this family. Direct sequencing of the PCR product from the patient II-2 is shown on the right. Arrow indicates the 912 C>A change.
Mentions: We initially screened PCR products of all seven exons of the CTSC gene for mutation(s) in one patient from each family using DNA sequencing technique (Fig. 2). Once the mutation was detected, it was confirmed in other members of the family. Sequence analysis of the exon 1 PCR product from the patient III-1 of the family IISC-PLS1 showed a nucleotide change in a homozygous state at position 145 from C>T (c.145C>T) which introduced a premature stop codon in the exon 1 (p.Q49X) (Fig. 2A). This change was also present in affected sibling III-2 in a homozygous state, whereas both parents were heterozygous for this change (Fig. 2A). PCR-SSCP analysis showed that this change was not present in 60 normal chromosomes (data not shown).

Bottom Line: This study reported three novel nonsense mutations in three Indian families.These novel nonsense mutations are predicted to produce truncated dipeptidyl-peptidase I causing PLS phenotype in these families.A review of the literature along with three novel mutations reported here showed that the total number of mutations in the CTSC gene described to date is 41 with 17 mutations being located in exon 7.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Reproduction Development and Genetics, Indian Institute of Science, Bangalore, India. selvacell@rediffmail.com

ABSTRACT

Background: PLS is a rare autosomal recessive disorder characterized by early onset periodontopathia and palmar plantar keratosis. PLS is caused by mutations in the cathepsin C (CTSC) gene. Dipeptidyl-peptidase I encoded by the CTSC gene removes dipeptides from the amino-terminus of protein substrates and mainly plays an immune and inflammatory role. Several mutations have been reported in this gene in patients from several ethnic groups. We report here mutation analysis of the CTSC gene in three Indian families with PLS.

Methods: Peripheral blood samples were obtained from individuals belonging to three Indian families with PLS for genomic DNA isolation. Exon-specific intronic primers were used to amplify DNA samples from individuals. PCR products were subsequently sequenced to detect mutations. PCR-SCCP and ASOH analyses were used to determine if mutations were present in normal control individuals.

Results: All patients from three families had a classic PLS phenotype, which included palmoplantar keratosis and early-onset severe periodontitis. Sequence analysis of the CTSC gene showed three novel nonsense mutations (viz., p.Q49X, p.Q69X and p.Y304X) in homozygous state in affected individuals from these Indian families.

Conclusions: This study reported three novel nonsense mutations in three Indian families. These novel nonsense mutations are predicted to produce truncated dipeptidyl-peptidase I causing PLS phenotype in these families. A review of the literature along with three novel mutations reported here showed that the total number of mutations in the CTSC gene described to date is 41 with 17 mutations being located in exon 7.

Show MeSH
Related in: MedlinePlus