Cell type-specific roles of histone deacetylase in TR ligand-independent transcriptional repression.
Recent evidence indicates that corepressor protein with histone deacetylase (HDAC) activity mediates thyroid hormone receptor (TR) transcriptional repression. In order to examine the physiological relevance of HDAC in ligand-independent TR-mediated repression, we studied the effect of trichostatin A (TSA), a specific HDAC inhibitor, in transient transfection studies with natural reporters, and assessed the expression of TR-regulated endogenous genes. Luciferase-coupled DR4-, F2-, PAL- or GH-TREs and TRbeta1 expression vectors were cotransfected in CV-1 and GH(3) cells. We did not observe any effect of TSA on TR-induced basal repression in CV-1 cells. Instead, TSA was able to induce an increase in transcription without T(3) on all TREs tested in GH(3) cells. This increase was >7-fold on F2-, >4-fold on DR4-, and 3-fold on GH-TREs. The cotransfection of a TRbeta1 mutant that exhibits decreased affinity with N-CoR (AHT) reduced the TSA effect in GH(3) cells, demonstrating a primary role for TR/N-CoR/Sin3/HDAC complex. Next, we examined the effects of TSA on endogenous GH mRNA production in GH(3) cells by Northern blot analysis. We observed an increase of 50-70% of GH mRNA in cells treated with TSA in hypothyroid medium, and an increase of GH mRNA in T(3)-treated cells after TSA treatment. Our results show that TSA can increase the expression of endogenous genes that are susceptible to TR regulation. These results support an active role of HDAC in transcriptional repression by ligand-independent TR.
Lizcano
F
F
Division of Genetics, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA. flizcano@rics.bwh.harvard.edu
Koibuchi
N
N
Fukuda
H
H
Dangond
F
F
Chin
W W
WW
D000818:Animals; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D013006:Growth Hormone / Q000235:genetics; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000235:genetics* / Q000502:physiology; D006877:Hydroxamic Acids / Q000494:pharmacology; D007668:Kidney / Q000166:cytology; D009928:Organ Specificity; D010902:Pituitary Gland / Q000166:cytology; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000235:genetics*; D012097:Repressor Proteins / Q000494:pharmacology*; D020218:Response Elements / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D011988:Receptors, Thyroid Hormone; D012097:Repressor Proteins; C012589:trichostatin A; D013006:Growth Hormone; D006655:Histone Deacetylases