| Project/Area Number |
05670718
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | HIROSAKI UNIVERSITY |
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Principal Investigator |
SAWAMURA Daisuke Hirosaki Univ.School of Med, Dermatology, assistant professor, 医学部附属病院, 講師 (60196334)
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| Co-Investigator(Kenkyū-buntansha) |
TAMAI Katsuto Hirosaki Univ.School of Med, Dermatology, assistant professor, 医学部附属病院, 講師 (20236730)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Bullous pemphigoid antigens(BPA) / Bullous pemphigoid / 5 prime flanking region / Keratinocyte / Keratinocyte specific expressio / Cis-acting element / 5prime領域 / ケラチノサイト特異発現 / 自己免疫性水疱症 / ヘミデスモゾーム / 基底膜 / 調節領域 / 調節エレメント / 類点疱瘡抗原 |
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| Research Abstract |
Bullous pemphigoid antigens (BPA) are proteins which are recognized by circulating autoantibodies in the sera of the patients with bullous pemphigoid. At least two antigens have been reported so far ; one of which is a 230 kDa protein, wereas another protein is 180 kDa collagen. This study intended to characterize the 5 prime flanking region of 230 kDa BPA gene in order to know regulation of this gene expression. First, we screened a human and a mouse genomic libraries with a human 230 kDa BPA cDNA,resulting in human and mouse positive clones. Nucleotide sequencing of these clones showed that some clones contained the 5 prime flanking regions. And the sequences of about 3 kb of 5 prime flanking regions detected several putative cis-acting elements. High sequence homology between the human and mouse 5 prime flanking regions suggested that regulation of this gene expression is critical for the function of keratinocytes. Furthermore, after PCR amplification of a human 2kb segment and of a mouse 1.2kb segment upstream from translation starting sites, we made chlolamphenicol acetyltranferase (CAT) constructs with these fragments, and transfected them to cultured cells including keratinocytes. Detection of CAT activity in only keratinocytes indicated that these fragments contained cis-elements responsible for keratinocyte specific expression. We constructed deletion mutants and transfected them to keratinocytes to examine localization of the elements. The CAT activities revieled that the elements for keratinocyte specific expression resided in -216 to -197 of the human 5 prime flanking region, and in -525 to -213 of the mouse region.
We made a few studies accompanied with the study mentioned above. We showed chromosomal localization of mouse 230 kDa BPA gene using the mouse clone, and examined a role of AP-1 site in the promoter of stromelysin gene in UV-induced skin changes to clarify the function of the AP-1 site in 5 prime flanking region of 230 kDa BPA gene.
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