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类LCR的α-珠蛋白阳性调控元件在红系分化过程中，在瞬时转染细胞中作为增强子发挥作用。

The LCR-like alpha-globin positive regulatory element functions as an enhancer in transiently transfected cells during erythroid differentiation.

作者信息

Pondel M D, George M, Proudfoot N J

机构信息

Sir William Dunn School of Pathology, University of Oxford, UK.

出版信息

Nucleic Acids Res. 1992 Jan 25;20(2):237-43. doi: 10.1093/nar/20.2.237.

DOI:10.1093/nar/20.2.237
PMID:1741249
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC310360/
Abstract

A positive regulatory element (PRE) similar to the locus control region (LCR) of the human beta-globin gene cluster has recently been identified 40 kb upstream of the human zeta-globin mRNA cap site (Higgs D.R. W.G. Wood, A.P. Jarman, J. Sharpe, J. Lida, I.M. Pretorius, and H. Ayyub. 1990). We investigated the influence of the alpha PRE on human alpha-globin promoter activity in transiently transfected cells. The introduction of the alpha PRE into alpha-globin promoter/CAT expression constructs increased alpha-globin promoter activity by 15-30 fold in a human erythroid cell line (Putko) as well as in mouse erythroleukemia cells (MELCs) induced with hexamethylene bisacetamide (HMBA). When these constructs were introduced into uninduced MELCs or HeLa cells, only a 2-3 fold increase in alpha-globin promoter activity was observed. Deletion of 600 bp of alpha-globin 5' flanking sequences containing six putative SP1-binding sites had no significant effect on levels of alpha-globin promoter enhancement by the alpha PRE. We further demonstrated that the alpha PRE and HS2 of the beta-LCR could similarly enhance transcriptional activity of the SV40 early promoter in HMBA induced MELCs. Finally, we showed that alpha-globin promoter activity in the presence of the alpha PRE increased with continued HMBA exposure and was coincident with transcriptional activation of endogenous globin genes.

摘要

最近，在人ζ-珠蛋白mRNA帽位点上游40 kb处发现了一种类似于人β-珠蛋白基因簇基因座控制区（LCR）的正向调控元件（PRE）（希格斯·D.R.、W.G.伍德、A.P.贾曼、J.夏普、J.利达、I.M.普雷托里乌斯和H.阿尤布，1990年）。我们研究了α-PRE对瞬时转染细胞中人α-珠蛋白启动子活性的影响。将α-PRE引入α-珠蛋白启动子/CAT表达构建体后，在人红系细胞系（普特科细胞）以及用六亚甲基双乙酰胺（HMBA）诱导的小鼠红白血病细胞（MELCs）中，α-珠蛋白启动子活性增加了15至30倍。当将这些构建体引入未诱导的MELCs或HeLa细胞时，仅观察到α-珠蛋白启动子活性增加了2至3倍。缺失包含六个假定SP1结合位点的600 bpα-珠蛋白5'侧翼序列，对α-PRE增强α-珠蛋白启动子的水平没有显著影响。我们进一步证明，α-PRE和β-LCR的HS2在HMBA诱导的MELCs中可同样增强SV40早期启动子的转录活性。最后，我们表明，在存在α-PRE的情况下，α-珠蛋白启动子活性随着持续暴露于HMBA而增加，并且与内源性珠蛋白基因的转录激活同时发生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/fd157cadb79f/nar00076-0077-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/c9f58cbc3fb4/nar00076-0075-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/2c32650f4e11/nar00076-0075-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/ec5dedb40682/nar00076-0076-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/fd157cadb79f/nar00076-0077-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/c9f58cbc3fb4/nar00076-0075-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/2c32650f4e11/nar00076-0075-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/ec5dedb40682/nar00076-0076-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ef20/310360/fd157cadb79f/nar00076-0077-a.jpg

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