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多瘤病毒增强子在整合到次黄嘌呤磷酸核糖基转移酶基因时可激活染色质可及性。

The polyomavirus enhancer activates chromatin accessibility on integration into the HPRT gene.

作者信息

Pikaart M, Feng J, Villeponteau B

机构信息

Department of Biological Chemistry, University of Michigan, Ann Arbor 48109-2007.

出版信息

Mol Cell Biol. 1992 Dec;12(12):5785-92. doi: 10.1128/mcb.12.12.5785-5792.1992.

DOI:10.1128/mcb.12.12.5785-5792.1992
PMID:1333045
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC360518/
Abstract

Recent studies suggest that enhancers may increase the accessibility of chromatin to transcription factors. To test the effects of a viral enhancer on chromatin accessibility, we have inserted minigenes with or without the polyomavirus enhancer into the third exon of the hypoxanthine phosphoribosyltransferase (HPRT) gene by homologous recombination and have prepared high-resolution maps of gene accessibility by using a novel polymerase chain reaction assay for DNase I sensitivity. In its native state, we find that the HPRT gene has low sensitivity to DNase I in fibrosarcoma cells. Insertion of the polyomavirus enhancer and neo reporter gene into exon 3 confers altered HPRT DNase I sensitivity for several kilobases on either side of the enhancer. The changes in DNase I sensitivity peak near the enhancer and decline with distance from the enhancer. The increase in HPRT DNase I sensitivity persisted when the tk promoter was deleted from the inserted construct but disappeared when the enhancer was deleted. These experiments identify the polyomavirus enhancer as a cis-acting initiator of chromatin accessibility.

摘要

近期研究表明，增强子可能会增加染色质对转录因子的可及性。为了测试病毒增强子对染色质可及性的影响，我们通过同源重组将含有或不含有多瘤病毒增强子的微型基因插入到次黄嘌呤磷酸核糖转移酶（HPRT）基因的第三个外显子中，并使用一种针对DNase I敏感性的新型聚合酶链反应分析法制备了基因可及性的高分辨率图谱。在其天然状态下，我们发现HPRT基因在纤维肉瘤细胞中对DNase I的敏感性较低。将多瘤病毒增强子和新霉素报告基因插入到外显子3中，会使增强子两侧数千碱基的HPRT对DNase I的敏感性发生改变。DNase I敏感性的变化在增强子附近达到峰值，并随着与增强子距离的增加而下降。当从插入的构建体中删除tk启动子时，HPRT对DNase I敏感性的增加仍然存在，但当增强子被删除时则消失。这些实验确定多瘤病毒增强子是染色质可及性的顺式作用引发剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/37829feb7047/molcellb00135-0520-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/509d1edeb784/molcellb00135-0517-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/deca6526a2ae/molcellb00135-0518-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/b5f8afbbc2f9/molcellb00135-0519-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/37829feb7047/molcellb00135-0520-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/509d1edeb784/molcellb00135-0517-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/deca6526a2ae/molcellb00135-0518-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/b5f8afbbc2f9/molcellb00135-0519-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/81b5/360518/37829feb7047/molcellb00135-0520-a.jpg

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