Two different transcription factors discriminate the -315C>T polymorphism of the Fc epsilon RI alpha gene: binding of Sp1 to -315C and of a high mobility group-related molecule to -315T.
The alpha-chain is a specific component of FcepsilonRI, which is essential for the cell surface expression of FcepsilonRI and the binding of IgE. Recently, two single nucleotide polymorphisms (SNPs) in the alpha-chain promoter, -315C>T and -66T>C, have been shown by statistic studies to associate with allergic diseases. The effect of -66 SNP on GATA-1-mediated promoter activity has been already indicated. In the present study, to investigate roles of the -315 SNP on the alpha-chain promoter functions, the transcription activity was evaluated by reporter assay. The alpha-chain promoter carrying -315T (minor allele) possessed significantly higher transcriptional activity than that of -315C (major allele). EMSA indicated that the transcription factor Sp1, but not Myc-associated zinc finger protein (MAZ), was bound to the -315C allele probe and that a transcription factor belonging to a high mobility group-family bound to the -315T allele probe. The chromatin immunoprecipitation assay suggested that high mobility group 1, 2, and Sp1 bound around -315 of FcepsilonRIalpha genomic DNA in vivo in the human basophil cell line KU812 with -315C/T and in human peripheral blood basophils with -315C/C, respectively. When cell surface expression level of FcepsilonRI on basophils was analyzed by flow cytometry, basophils from individuals carrying -315T allele expressed significantly higher amount of FcepsilonRI compared with those of -315C/C. The findings demonstrate that a -315 SNP significantly affects human FcepsilonRI alpha-chain promoter activity and expression level of FcepsilonRI on basophils by binding different transcription factors to the SNP site.
Kanada
Shunsuke
S
Atopy (Allergy) Research Center, Juntendo University School of Medicine, Tokyo, Japan.
Nakano
Nobuhiro
N
Potaczek
Daniel P
DP
Maeda
Keiko
K
Shimokawa
Naomi
N
Niwa
Yusuke
Y
Fukai
Tatsuo
T
Sanak
Marek
M
Szczeklik
Andrew
A
Yagita
Hideo
H
Okumura
Ko
K
Ogawa
Hideoki
H
Nishiyama
Chiharu
C
D000483:Alleles; D000818:Animals; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003597:Cytosine Nucleotides / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide / Q000276:immunology*; D011401:Promoter Regions, Genetic / Q000276:immunology; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D017455:Receptors, IgE / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D003597:Cytosine Nucleotides; D004268:DNA-Binding Proteins; C475664:FCER1A protein, human; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; D021122:Protein Subunits; D017455:Receptors, IgE; D016329:Sp1 Transcription Factor; D013942:Thymine Nucleotides; D014157:Transcription Factors; C076035:c-MYC-associated zinc finger protein