Genetic and molecular characterisation of a disintegrin and metalloprotease (ADAM) 33
Genetic and molecular characterisation of a disintegrin and metalloprotease (ADAM) 33
Polymorphism of the gene, ADAM33, is associated with increased susceptibility to asthma and bronchial hyperresponsiveness (BHR). This work investigates the association of ADAM33 polymorphism with early-life lung function and characterises basal transcriptional activation of ADAM33.
5’ nuclease allelic discrimination assays were developed and used to genotype a one month old cohort with infant lung function data (COPSAC, n=401). Haplotype trend regression, haploscore analyses and linkages disequilibrium mapping were used to assess association with lung function. Transcriptional regulation of ADAM33 was investigated using deletion analysis of ~1500 bp of putative basal promoter sequence. Sequences were cloned into pG13 basic vectors and Dual-Luciferase® reporter assays used to assess relative activity. Cross species and intra-species analysis was performed, using Vista, to identify conserved regions of the ADAM33 promoter. Putative transcription binding sites were predicted using MatInspector.
ADAM33 polymorphisms was not associated with lung function at four weeks of age, contrasting with a previous study in a population of 3 year olds. Dual-Luciferase® reporter assays suggested the core promoter region lies within 359 bp (F3X) of the translation start site (ATG) and there is an inhibitory site between -637 bp and -359 bp. MatInspector predicted Sp1 sites within the F3X sequence. Over expression with Sp1 reduced ADAM33 expression in fibroblast cell lines, and increased expression in epithelial cell lines indicating an activation mechanism involving Sp1.
This work implicates Sp1 as a key regulator of ADAM33 transcription. Polymorphic changes were not associated with reduced lung function at one month old, suggesting a window of opportunity exists between birth and three years in which gene-environmental interactions take place to trigger asthmatic symptoms in an already pre-disposed lung.
University of Southampton
Cakebread, Julie Ann
6fcff755-c817-441a-9f9d-e7ea5f8e793d
2006
Cakebread, Julie Ann
6fcff755-c817-441a-9f9d-e7ea5f8e793d
Cakebread, Julie Ann
(2006)
Genetic and molecular characterisation of a disintegrin and metalloprotease (ADAM) 33.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Polymorphism of the gene, ADAM33, is associated with increased susceptibility to asthma and bronchial hyperresponsiveness (BHR). This work investigates the association of ADAM33 polymorphism with early-life lung function and characterises basal transcriptional activation of ADAM33.
5’ nuclease allelic discrimination assays were developed and used to genotype a one month old cohort with infant lung function data (COPSAC, n=401). Haplotype trend regression, haploscore analyses and linkages disequilibrium mapping were used to assess association with lung function. Transcriptional regulation of ADAM33 was investigated using deletion analysis of ~1500 bp of putative basal promoter sequence. Sequences were cloned into pG13 basic vectors and Dual-Luciferase® reporter assays used to assess relative activity. Cross species and intra-species analysis was performed, using Vista, to identify conserved regions of the ADAM33 promoter. Putative transcription binding sites were predicted using MatInspector.
ADAM33 polymorphisms was not associated with lung function at four weeks of age, contrasting with a previous study in a population of 3 year olds. Dual-Luciferase® reporter assays suggested the core promoter region lies within 359 bp (F3X) of the translation start site (ATG) and there is an inhibitory site between -637 bp and -359 bp. MatInspector predicted Sp1 sites within the F3X sequence. Over expression with Sp1 reduced ADAM33 expression in fibroblast cell lines, and increased expression in epithelial cell lines indicating an activation mechanism involving Sp1.
This work implicates Sp1 as a key regulator of ADAM33 transcription. Polymorphic changes were not associated with reduced lung function at one month old, suggesting a window of opportunity exists between birth and three years in which gene-environmental interactions take place to trigger asthmatic symptoms in an already pre-disposed lung.
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Published date: 2006
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Local EPrints ID: 466065
URI: http://eprints.soton.ac.uk/id/eprint/466065
PURE UUID: d9cfc1fc-c5a4-47d6-8959-65826c291843
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Date deposited: 05 Jul 2022 04:13
Last modified: 16 Mar 2024 20:29
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Author:
Julie Ann Cakebread
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