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维生素D反应性骨钙素转录单位在酿酒酵母中的重建。

Reconstitution of the vitamin D-responsive osteocalcin transcription unit in Saccharomyces cerevisiae.

作者信息

McDonnell D P, Pike J W, Drutz D J, Butt T R, O'Malley B W

机构信息

Department of Anti-Infectives, Smith Kline and French Laboratories, King of Prussia, Pennsylvania 19406.

出版信息

Mol Cell Biol. 1989 Aug;9(8):3517-23. doi: 10.1128/mcb.9.8.3517-3523.1989.

DOI:10.1128/mcb.9.8.3517-3523.1989
PMID:2552296
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC362399/
Abstract

The human osteocalcin gene is regulated in mammalian osteoblasts by 1,25(OH)2D3-dependent and -independent mechanisms. The sequences responsible for this activity have been mapped to within the -1339 region of the gene. We show here that this enhancer region functions analogously in Saccharomyces cerevisiae cells engineered to produce active 1,25(OH)2D3 receptor. When fused to the proximal promoter elements of the yeast iso-1-cytochrome c gene, the enhancer demonstrated substantial promoter activity. This activity was elevated further by 1,25(OH)2D3 when the reporter constructs were assayed in cells containing the 1,25(OH)2D3 receptor. This system affords a model for 1,25(OH)2D3 action and represents a simple assay system that will enable definition of the important cis-acting regulatory sequences within the osteocalcin gene and identification of their cognate transcription factors.

摘要

人骨钙素基因在哺乳动物成骨细胞中受1,25(OH)₂D₃依赖性和非依赖性机制调控。负责此活性的序列已定位到该基因的-1339区域内。我们在此表明，该增强子区域在经工程改造以产生活性1,25(OH)₂D₃受体的酿酒酵母细胞中具有类似功能。当与酵母异-1-细胞色素c基因的近端启动子元件融合时，该增强子表现出显著的启动子活性。当在含有1,25(OH)₂D₃受体的细胞中检测报告构建体时，1,25(OH)₂D₃可进一步提高这种活性。该系统为1,25(OH)₂D₃的作用提供了一个模型，并且代表了一个简单的检测系统，能够确定骨钙素基因内重要的顺式作用调控序列并鉴定其同源转录因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa8a/362399/f7e4993d3c3d/molcellb00056-0375-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa8a/362399/f7e4993d3c3d/molcellb00056-0375-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa8a/362399/f7e4993d3c3d/molcellb00056-0375-a.jpg

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A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site.一个GAL10-CYC1杂交酵母启动子将GAL4调控区域鉴定为一个上游位点。
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