Promoter activity of human tissue inhibitor of metalloproteinase 2 gene with novel single nucleotide polymorphisms.
OBJECTIVE
The single nucleotide polymorphism (SNP) -418G > C in the TIMP2 gene promoter region has been shown to be associated with in chronic obstructive pulmonary disease (COPD). The purpose of this study was to search for novel single nucleotide polymorphism (SNP) in the TIMP2 promoter region around the -418G > C locus, and to investigate whether any of these SNP, including -418G > C, had an influence on TIMP2 transcription activity.


METHODS
DNA sequencing was performed on a 689 base-pair polymerase chain reaction fragment of the promoter region. The novel SNP were characterized and genotype analysis was performed for COPD and control subjects. A reporter gene assay was performed using the wild-type promoter (-418G/-177C/+34C) and the mutant-type promoter (-418C/-177T/+34A).


RESULTS
Nine novel SNP were identified. The SNP -177C > T and +34C > A were in complete linkage disequilibrium with -418G > C. The other seven SNP were not associated with COPD. No significant difference was detected in the reporter gene assay between the activities of the wild-type and the mutant-type promoters.


CONCLUSIONS
The SNP -418G > C, -177C > T and +34C > A, might not themselves be functional from a transcriptional point of view in the development of COPD, but may be in linkage disequilibrium with other functional polymorphisms.
Hegab
Ahmed E
AE
Department of Pulmonary Medicine, Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan.
Sakamoto
Tohru
T
Uchida
Yoshiyuki
Y
Nomura
Akihiro
A
Ishii
Yukio
Y
Morishima
Yuko
Y
Mochizuki
Mie
M
Kimura
Toru
T
Saitoh
Wataru
W
Iizuka
Takashi
T
Kiwamoto
Takumi
T
Sekizawa
Kiyohisa
K
D020029:Base Pairing / Q000235:genetics; D016022:Case-Control Studies; D003596:Cytosine; D005787:Gene Frequency / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D008040:Genetic Linkage / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D005838:Genotype; D006147:Guanine; D006801:Humans; D009154:Mutation / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D029424:Pulmonary Disease, Chronic Obstructive / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection
D019716:Tissue Inhibitor of Metalloproteinase-2; D006147:Guanine; D003596:Cytosine