An insulator element 3' to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells. Export

@article{citeulike:4129019, abstract = {Regulation of expression of the CFTR gene is poorly understood. Elements within the basal promoter of the gene do not fully explain CFTR expression patterns, suggesting that cis-regulatory elements are located elsewhere, either within the locus or in adjacent chromatin. We previously mapped DNase I hypersensitive sites (DHS) in 400 kb spanning the CFTR locus including a cluster of sites close to the 3'-end of the gene. Here we focus on a DHS at +6.8 kb from the CFTR translation end-point to evaluate its potential role in regulating expression of the gene. This DHS, which encompasses a consensus CTCF-binding site, was evident in primary human epididymis cells that express abundant CFTR mRNA. We show by DNase I footprinting and electophoretic mobility shift assays that the cis-regulatory element within this DHS binds CTCF in vitro. We further demonstrate that the element functions as an enhancer blocker in a well-established in vivo assay, and by using chromatin immunoprecipitation that it recruits CTCF in vivo. Moreover, we reveal that in primary epididymis cells, the +6.8 kb DHS interacts closely with the CFTR promoter, suggesting that the CFTR locus exists in a looped conformation, characteristic of an active chromatin hub.}, author = {Blackledge, Neil P P. and Ott, Christopher J J. and Gillen, Austin E E. and Harris, Ann}, citeulike-article-id = {4129019}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/19129223}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=19129223}, day = {12}, issn = {1362-4962}, journal = {Nucleic acids research}, keywords = {3, cftr, ctcf, insulator}, month = {January}, posted-at = {2009-03-03 19:06:40}, priority = {5}, title = {An insulator element 3' to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/19129223}, year = {2009} }

TY - JOUR ID - citeulike:4129019 L3 - citeulike-article-id:4129019 N2 - Regulation of expression of the CFTR gene is poorly understood. Elements within the basal promoter of the gene do not fully explain CFTR expression patterns, suggesting that cis-regulatory elements are located elsewhere, either within the locus or in adjacent chromatin. We previously mapped DNase I hypersensitive sites (DHS) in 400 kb spanning the CFTR locus including a cluster of sites close to the 3'-end of the gene. Here we focus on a DHS at +6.8 kb from the CFTR translation end-point to evaluate its potential role in regulating expression of the gene. This DHS, which encompasses a consensus CTCF-binding site, was evident in primary human epididymis cells that express abundant CFTR mRNA. We show by DNase I footprinting and electophoretic mobility shift assays that the cis-regulatory element within this DHS binds CTCF in vitro. We further demonstrate that the element functions as an enhancer blocker in a well-established in vivo assay, and by using chromatin immunoprecipitation that it recruits CTCF in vivo. Moreover, we reveal that in primary epididymis cells, the +6.8 kb DHS interacts closely with the CFTR promoter, suggesting that the CFTR locus exists in a looped conformation, characteristic of an active chromatin hub. JF - Nucleic acids research SN - 1362-4962 TI - An insulator element 3' to the CFTR gene binds CTCF and reveals an active chromatin hub in primary cells. KW - 3 KW - cftr KW - ctcf KW - insulator AU - Blackledge, Neil P AU - Ott, Christopher J AU - Gillen, Austin E AU - Harris, Ann PY - 2009/01/12/ UR - http://view.ncbi.nlm.nih.gov/pubmed/19129223 ER -