title: Detection of a large genomic deletion in the pancreatic secretory trypsin inhibitor (SPINK1) gene.
abstract: Mutations and polymorphisms in the SPINK1 gene, which encodes trypsin's physiological inhibitor, pancreatic secretory trypsin inhibitor, have been found to be associated with chronic pancreatitis. However, to date, all currently reported SPINK1 variants are either single-nucleotide substitutions or microinsertions/deletions. It is possible that large genomic rearrangements at this locus may underlie certain cases of chronic pancreatitis. However, such events, if indeed they exist, may have been overlooked by conventional PCR-based techniques. Here we attempted to screen all four exons as well as the promoter region of the SPINK1 gene for large genomic deletions by means of quantitative high-performance liquid chromatography analysis. Of the 47 pancreatitis families (not carrying any known PRSS1, SPINK1 and CFTR variants/mutations after screening the coding regions by our previously established denaturing high-performance liquid chromatography methods), one family was suggested to carry a large genomic deletion in the SPINK1 gene. The aberrant chromosomal junction was encapsulated by long-range PCR and the breakpoints were determined by direct sequencing of the rearranged fragment. A 2-bp short direct repeat was present at the deletion breakpoints; this simple deletion (c.1-320_c.55+961del1336 bp) can thus in principle be explained by replication slippage. Identification of this lesion has not only expanded the SPINK1 mutational spectrum but also served to identify a novel mutational mechanism causing chronic pancreatitis.
authors.affiliation: INSERM, U613, 29220 Brest, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Masson
authors.forename: Cédric
authors.initials: C
authors.lastname: Le Maréchal
authors.forename: Jian-Min
authors.initials: JM
authors.lastname: Chen
authors.forename: Thierry
authors.initials: T
authors.lastname: Frebourg
authors.forename: Eric
authors.initials: E
authors.lastname: Lerebours
authors.forename: Claude
authors.initials: C
authors.lastname: Férec
mesh_terms: D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D005091:Exons; D005260:Female; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D050500:Pancreatitis, Chronic / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014359:Trypsin Inhibitor, Kazal Pancreatic
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; C486642:SPINK1 protein, human; D014359:Trypsin Inhibitor, Kazal Pancreatic