Transcriptional control mechanism for the expression of type I collagen gene in fibrotic skin disorders and related diseases

纤维化皮肤病及相关疾病中 I 型胶原基因表达的转录控制机制

• 批准号: 08670948
• 负责人: HATAMOCHI Atsushi
• 金额: $ 1.41万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670948/
• 关键词: type I collagen; promoter gene; Pur alpha; Pur beta; TGF beta; TGF beta receptor; transcription; Purα; Purβ; TGFβ; TGFβレセプター; 遺伝子発現; TNF-α; COLF1; 皮膚線維症

Collagen type I,a most abundant protein in the dermis, consists of two alphal(I) chain and one alpha2(I) chain which are coordinately expressed. ColFl, a DNA binding protein which specifically binds to a segment of the alpha2(I)collagen promoter at -400bp upstream of the start of transcription, activates transcription of the alpha2(I)collagen gene in vitro. We investigated on the partial sequences of polypeptide and the cDNA cloning of this transcriptional factor of the alpha2(I)collagen gene. The protein purified by sequence-specific DNA affinity chromatography were found to consist of 42kDa and 40.5 kDa polypeptides. Sequences of peptide fragments from 42kDa polypeptide were identical to Pur alpha, is a nuclear protein which has been reported to binds to a upstream region of human c-myc gene. Some of those from 40kDa polypeptide were identical to Pur beta, has been partially sequenced and has regions of strong homology to Pur alpha Full length of Pur beta cDNA were sequenced. The deductive amino acid sequences of Pur alpha and Pur beta showed 61.4% homology. Previous studies have demonstrated that the expression of type I collagen, the most abundant protein in the dermis, is reduced in in vitro aging fibroblast cultures, but the mechanism controlling the reduction of type I collagen expression is not understood. We found that the mRNA levels of alphal(I) collagen, TGFbeta, and TGFbeta receptors I and II in late-passage fibroblasts were reduced, and the TGF beta receptor binding in late-passage fibroblasts was lower than that in early-passage fibroblasts.

I型胶原是真皮中最丰富的蛋白质，由两条α 1（I）链和一条α 2（I）链组成，它们协同表达。ColFl是一种DNA结合蛋白，其特异性结合于转录起始点上游约400 bp处的α 2（I）胶原启动子片段，在体外激活α 2（I）胶原基因的转录。本研究对α_2（I）胶原基因的部分多肽序列进行了测定，并克隆了该转录因子的cDNA。序列特异性DNA亲和层析纯化的蛋白质由42 kDa和40.5kDa的多肽组成。42 kDa多肽的肽段序列与Pur α相同，Pur α是已报道的与人c-myc基因上游区结合的核蛋白。其中一些来自40 kDa多肽，与Pur β相同，已部分测序，并具有与Pur α强同源性的区域。推导的Pur α和Pur β氨基酸序列同源性为61.4%。以前的研究已经证明，I型胶原蛋白，在真皮中最丰富的蛋白质，在体外老化的成纤维细胞培养物中的表达减少，但控制I型胶原蛋白表达减少的机制尚不清楚。我们发现，在晚期传代成纤维细胞中，胶原（I）、TGF β和TGF β受体I和II的mRNA水平降低，并且晚期传代成纤维细胞中的TGF β受体结合低于早期传代成纤维细胞。

项目成果

Mori Y, Hatamochi A, Arakawa M, Ueki H: "Reduced expression of mRNA for TGFβand TGFβ receptor I and II and decreased TGFβ binding in in vitro aged fibroblas" Arch Dermatol Res. (発表予定).

Mori Y、Hatamochi A、Arakawa M、Ueki H：“TGFβ 和 TGFβ 受体 I 和 II 的 mRNA 表达减少，并减少体外老化成纤维细胞中 TGFβ 的结合”Arch Dermatol Res。

Mori Y,Hatamochi A,Arakawa M,Ueki H: "Reduced expression of mRNA for transforming growth factor beta(TGFbeta) and TGFbeta receptors I and II and decreased TGFbeta binding to the receptors in in vitro aged fibroblasts" Arch Dermatol Res. (in press).

Mori Y、Hatamochi A、Arakawa M、Ueki H：“在体外老化成纤维细胞中，转化生长因子 β (TGFbeta) 和 TGFbeta 受体 I 和 II 的 mRNA 表达减少，并减少 TGFbeta 与受体的结合”Arch Dermatol Res。

Mori Y, Hatamochi A, Arakawa M, Ueki H: "Reduced expression of mRNA for transforming growth factor β and TGF β receptor I and II anddecreased TGF β binding in in vitro aged fibroblasts" Arch Dermatol Res. (発表予定).

Mori Y、Hatamochi A、Arakawa M、Ueki H：“转化生长因子 β 和 TGF β 受体 I 和 II 的 mRNA 表达减少，并减少体外老化成纤维细胞中 TGF β 的结合”Arch Dermatol Res。

Kenichi More: "The transcription of human α1(I) procollagen gene (COL1A1) is suppressed by tumor necrosis factor-α through proximal short promoter elements" Biochem.J.319. 811-816 (1996)

Kenichi More：“肿瘤坏死因子-α 通过近端短启动子元件抑制人 α1(I) 前胶原基因 (COL1A1) 的转录”Biochem.J.319 (1996)。

籏持淳: "サイトカインとI型コラーゲン遺伝子の転写調節" connective Tissue. 28. 197-204 (1996)

Jun Momochi：“细胞因子和 I 型胶原蛋白基因的转录调控”结缔组织 28. 197-204 (1996)。