REGULATION OF COLLAGEN EXPRESSION IN CHONDROCYTES

软骨细胞中胶原蛋白表达的调节

• 批准号: 3319083
• 负责人: HELGA B DOTY
• 金额: $ 7.64万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-08-01 至 1988-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3319083
• 关键词: DNA; DNA methylation; cartilage development; cell differentiation; cell transformation; chondrocytes; chromatin; deoxyribonuclease I; early embryonic stage; fibroblasts; genetic manipulation; genetic transcription; genetic translation; histogenesis; limbs; molecular cloning; nonmammalian vertebrate embryology; nucleic acid sequence; plasmids; regulatory gene; structural genes

The proposed research is directed at identifying DNA sequences, both within and surrounding the type I and type II chicken collagen genes, which are involved in either positive or negative regulation of these genes in differentiated fibroblasts and chondrocytes. The state of methylation of both type I and type II collagen genes will be reinvestigated using a modified gel transfer procedure which will permit detection of very small fragments. Changes in chromatin structure, seen as DNAse I hypersensitive sites, will be looked for in type I and type II collagen genes and flanking gene regions in both fibroblast and chondrocyte nuclei to determine whether changes, if they occur, are associated with expression. To identify sequences involved in regulation fibroblasts and chondrocytes will be transiently transformed with plasmids containing from 2000 to less than 100 bp of 5' flanking gene sequences of the pro Alpha1(I), pro Alpha2(I) and pro Alpha1(II) collagen genes placed 5' to genes whose expression can be readily tested at the protein and RNA level, such as SV40 large T and small t antigens. Analogous transient expression experiments will be carried out using collagen mimigenes in which the 5' flanking region and first three or more exons are ligated to part of the last intron, 3' most exon and 3' flanking gene region to determine if genomic regions and/or 3' flanking regions are involved in regulation. Once regulatory sequences are identified, it may be feasible to use them to isolate transacting factors involved in regulation. Obviously the failure to properly regulate the precisely timed sequential expression of type I and type II collagen genes would result in abnormal limb development.

拟议的研究是针对识别DNA序列，无论是在

项目成果

Construction and characterization of cDNA clones encoding the 5' end of the chicken pro alpha 1(I) collagen mRNA.

编码鸡 pro alpha 1(I) 胶原蛋白 mRNA 5 端的 cDNA 克隆的构建和表征。

• DOI: 10.1016/0378-1119(87)90159-4
• 发表时间: 1987
• 期刊: Gene
• 影响因子: 3.5
• 作者: Finer,MH;Boedtker,H;Doty,P
• 通讯作者: Doty,P

Unusual DNA sequences located within the promoter region and the first intron of the chicken pro-alpha 1(I) collagen gene.

不寻常的 DNA 序列位于鸡 pro-alpha 1(I) 胶原蛋白基因的启动子区域和第一个内含子内。

• 发表时间: 1987
• 期刊: The Journal of biological chemistry
• 作者: Finer,MH;Aho,S;Gerstenfeld,LC;Boedtker,H;Doty,P
• 通讯作者: Doty,P