Repression of Androgen Receptor Transcription through the E2F1/DNMT1 Axis
Publication Date
September 26, 2011
Journal
PLOS ONE
Authors
Conrad David Valdez, Joanne N. Davis, Hana M. Odeh, Tristan L. Layfield, et al
Volume
6
Issue
9
Pages
e25187
DOI
https://dx.plos.org/10.1371/journal.pone.0025187
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0025187
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/21966451
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3180375
Europe PMC
http://europepmc.org/abstract/MED/21966451
Web of Science
000295932100022
Scopus
80053142159
Mendeley
http://www.mendeley.com/research/repression-androgen-receptor-transcription-through-e2f1dnmt1-axis
Events
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Mendeley | Further Information

{"title"=>"Repression of androgen receptor transcription through the E2F1/DNMT1 axis", "type"=>"journal", "authors"=>[{"first_name"=>"Conrad David", "last_name"=>"Valdez", "scopus_author_id"=>"53265166900"}, {"first_name"=>"Joanne N.", "last_name"=>"Davis", "scopus_author_id"=>"57003877200"}, {"first_name"=>"Hana M.", "last_name"=>"Odeh", "scopus_author_id"=>"57191388078"}, {"first_name"=>"Tristan L.", "last_name"=>"Layfield", "scopus_author_id"=>"53264184000"}, {"first_name"=>"Craig S.", "last_name"=>"Cousineau", "scopus_author_id"=>"53263357800"}, {"first_name"=>"Thomas R.", "last_name"=>"Berton", "scopus_author_id"=>"6603164425"}, {"first_name"=>"David G.", "last_name"=>"Johnson", "scopus_author_id"=>"7406823034"}, {"first_name"=>"Kirk J.", "last_name"=>"Wojno", "scopus_author_id"=>"7004287350"}, {"first_name"=>"Mark L.", "last_name"=>"Day", "scopus_author_id"=>"7401703015"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"80053142159", "doi"=>"10.1371/journal.pone.0025187", "scopus"=>"2-s2.0-80053142159", "pui"=>"362627640", "pmid"=>"21966451"}, "id"=>"764c78f8-e997-33c4-a6eb-6cbb0c43a48e", "abstract"=>"Although androgen receptor (AR) function has been extensively studied, regulation of the AR gene itself has been much less characterized. In this study, we observed a dramatic reduction in the expression of androgen receptor mRNA and protein in hyperproliferative prostate epithelium of keratin 5 promoter driven E2F1 transgenic mice. To confirm an inhibitory function for E2F1 on AR transcription, we showed that E2F1 inhibited the transcription of endogenous AR mRNA, subsequent AR protein, and AR promoter activity in both human and mouse epithelial cells. E2F1 also inhibited androgen-stimulated activation of two AR target gene promoters. To elucidate the molecular mechanism of E2F-mediated inhibition of AR, we evaluated the effects of two functional E2F1 mutants on AR promoter activity and found that the transactivation domain appears to mediate E2F1 repression of the AR promoter. Because DNMT1 is a functional intermediate of E2F1 we examined DNMT1 function in AR repression. Repression of endogenous AR in normal human prostate epithelial cells was relieved by DNMT1 shRNA knock down. DNMT1 was shown to be physically associated within the AR minimal promoter located 22 bps from the transcription start site; however, methylation remained unchanged at the promoter regardless of DNMT1 expression. Taken together, our results suggest that DNMT1 operates either as a functional intermediary or in cooperation with E2F1 inhibiting AR gene expression in a methylation independent manner.", "link"=>"http://www.mendeley.com/research/repression-androgen-receptor-transcription-through-e2f1dnmt1-axis", "reader_count"=>10, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Unspecified"=>{"Unspecified"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/731622"], "description"=>"<p>(A) Map of a region of the AR promoter depicting two types of E2F consensus sequences +80→+96 and +13,318→+13,334 (shown as solid ovals) and +999→+1,015 (shown as open ovals). Primer flanked regions are designated as sites A, B, and C. The 243 bp region (+22→+293) analyzed by bisulfite sequencing is indicated. (B) qPCR analysis of target (DNMT1) and non-specific (IgG) ChIPed DNA from BPH-1 cells using primers that flank sites A, B, and C. Primers flanking a region in the PS2 (targeted DNMT1) promoter and ABCB1 (non-targeted DNMT1) region were used as ChIP controls. Data is representative of the mean from 3 qPCR reactions and shown as a percent of input with the standard error indicated. (C) Bisulfite sequencing analysis of the 243 bp region in the AR minimal promoter in hPrEC compared to DU145 cells infected with both non-targeting (NT) (control) and DNMT1 shRNA constructs (4-1 and 4-2). Solid circles (methylated) and open circles (un-methylated) were used to represent the methylation status of cytosines within CpG dinucleotides. Each horizontal strand of circles depicts a separate DNA clone. Lysates were probed on a western blot for AR and actin. Cell lines were also treated with either 1 µM 5Aza (5A) or DMSO matched vehicle (V) and extracted cDNA was PCR amplified with both human AR and GAPDH. All data shown except for the 5Aza treatments are representative of 3 separate experimental replicates.</p>", "links"=>[], "tags"=>["dnmt1", "ar"], "article_id"=>401969, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g005", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Methylation_independent_association_of_DNMT1_with_the_AR_gene_/401969", "title"=>"Methylation independent association of DNMT1 with the AR gene.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:25:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/731398"], "description"=>"<p>(<i>A)</i> Northern blot analysis of stably transfected mouse prostate epithelial cells (PrE) with either pCDNA3 (control vector) or E2F1 shown as E2F1-1 and E2F1-2 to detect AR and E2F1 gene transcription. The 28S and 18S ribosomal bands are shown for loading comparison. (<i>B)</i> Western blot analysis of whole cell lysates harvested from PrE cells stably transfected with pcDNA3 (control) or E2F1 for the detection of AR, Cyclin E, and PCNA. β-actin is shown as a loading control. (<i>A and B)</i> The northern blot and western is representative of 3 separate experiments. LNCaP cells were co-transfected with 1 µg of ARE-Luc (<i>C)</i> and 500 ng of either empty pcDNA3 vector or E2F1 in the presence of 10<sup>−9</sup> M R1881. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025187#s2\" target=\"_blank\">Results</a> were normalized to β-galactosidase (B-gal) from a co-transfected CMV promoter driven B-gal reporter construct. The histogram represents the mean value of three independent experiments with the indicated standard deviation. The western depicts the expression levels for E2F1 and AR relative to tubulin for the transfection and treatment conditions. * indicates P<0.05 for the indicated comparison in brackets.</p>", "links"=>[], "tags"=>["e2f1", "inhibits", "ar", "responsive"], "article_id"=>401747, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g002", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Exogenous_E2F1_inhibits_both_AR_expression_and_responsive_promoters_/401747", "title"=>"Exogenous E2F1 inhibits both AR expression and responsive promoters.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:24:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/731544"], "description"=>"<p>(A) Primary cultures of human prostate epithelial cells (hPrEC) and (B) immortalized benign prostate epithelial cells (BPH-1) transduced with either control (shVector (shV) and shNon-targeting (NT)) or DNMT1 shRNA constructs. AR and DNMT1 protein expression relative to actin loading control were analyzed by Western blot. The exposures in (B) were taken from different sections of the same blot at the same intensity. AR transcription was analyzed using qRTPCR with readings done in triplicate (graphs A and B). Mean values are represented with standard error bars. (C) shRNA transduced BPH-1 cells (described in A and B) were transfected with the human AR promoter luciferase reporter (hAR-Luc) construct. The histograms represent the mean value of three independent experiments with the indicated standard deviation. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025187#s2\" target=\"_blank\">Results</a> were normalized to β-galactosidase (B-gal) expression from a co-transfected CMV promoter driven B-gal reporter construct. All western blots are representative of 3 separate experimental replicates.</p>", "links"=>[], "tags"=>["downregulation", "relieves", "ar"], "article_id"=>401895, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g004", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DNMT1_downregulation_relieves_AR_Expression_/401895", "title"=>"DNMT1 downregulation relieves AR Expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:25:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/731322"], "description"=>"<p><i>(A)</i> Histology of prostate tissue taken from both K5-E2F1 transgenic and wild type mice. <i>(B)</i> Prostate epithelial cell lines established from both transgenic and wild type mice were analyzed by western blot for the expression of E2F1, cell cycle genes (Cyclin E and PCNA), the epithelial cell specific marker E-cadherin (E-cad), and Androgen Receptor (AR). Actin is shown as a loading control. <i>(C)</i> A trypan blue exclusion assay was implemented to measure the viability of cell lines obtained from the mouse models. Each point represents the mean of three independent experiments with the standard deviation.</p>", "links"=>[], "tags"=>["leads", "atypical", "prostatic", "morphology", "increases", "prostate", "epithelial", "proliferation", "k5-e2f1", "transgenic"], "article_id"=>401657, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g001", "stats"=>{"downloads"=>3, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_E2F1_leads_to_atypical_prostatic_morphology_and_increases_prostate_epithelial_proliferation_in_a_K5_E2F1_transgenic_mouse_/401657", "title"=>"E2F1 leads to atypical prostatic morphology and increases prostate epithelial proliferation in a K5-E2F1 transgenic mouse.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:23:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/731694"], "description"=>"<p>The use of either a dominant negative E2F1 construct or a shRNA to knock down DNMT1 both result in the downregulation of DNMT1 and subsequent rescue of AR expression. E2F1 overexpression experiments suggest that elevated E2F1 levels increase DNMT1 protein expression that associates directly with the AR promoter or possibly in complex with E2F1 to repress AR.</p>", "links"=>[], "tags"=>["ar", "repression"], "article_id"=>402045, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g006", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_AR_repression_through_the_E2F1_DNMT1_axis_/402045", "title"=>"Schematic representation of AR repression through the E2F1/DNMT1 axis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:25:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/731473"], "description"=>"<p>(<i>A)</i> mPrE cells were co-transfected with 1 µg of mARp-Luc, E2F-Luc or CRE-Luc luciferase reporter constructs with 0.5 µg of empty vector (pcDNA3), wild type E2F1, dominant negative E2F1 (DN E2F1) or SV-40 Large T antigen (Tag). After 72 hours, cells were harvested and assayed for luciferase expression. The results are shown as averages of three independent experiments. Assays were done in triplicate and mean values are shown with standard deviation. The results were normalized to β-galactosidase (B-gal) expression from a co-transfected CMV promoter driven B-gal reporter construct. <i>(B)</i> LNCaP cells were co-transfected with either 1 µg of hAR-Luc or DHFR-Luc and 0.5 µg of either wild type E2F1or E2F1<sub>1–284</sub> mutant constructs. The histograms represent the mean value of three independent experiments with the indicated standard deviation. <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0025187#s2\" target=\"_blank\">Results</a> were normalized to β-galactosidase (B-gal) expression from a co-transfected CMV promoter driven by a B-gal reporter construct. * indicates P<0.05 for the indicated comparison in brackets.</p>", "links"=>[], "tags"=>["e2f1", "transactivation", "ar", "promoter"], "article_id"=>401822, "categories"=>["Cancer", "Molecular Biology", "Genetics", "Developmental Biology"], "users"=>["Conrad David Valdez", "Joanne N. Davis", "Hana M. Odeh", "Tristan L. Layfield", "Craig S. Cousineau", "Thomas R. Berton", "David G. Johnson", "Kirk J. Wojno", "Mark L. Day"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0025187.g003", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_E2F1_transactivation_domain_is_required_for_AR_promoter_activity_repression_/401822", "title"=>"The E2F1 transactivation domain is required for AR promoter activity repression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 14:24:36"}

PMC Usage Stats | Further Information

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