Identification of Mouse Histone Deacetylase 1 as a Growth Factor-Inducible Gene

Abstract

Reversible acetylation of core histones plays an important role in transcriptional regulation, cell cycle progression, and developmental events. The acetylation state of histones is controlled by the activities of acetylating and deacetylating enzymes. By using differential mRNA display, we have identified a mouse histone deacetylase gene, HD1, as an interleukin-2-inducible gene in murine T cells. Sequence alignments revealed that murine HD1 is highly homologous to the yeast RPD3 pleiotropic transcriptional regulator. Indirect immunofluorescence microscopy proved that mouse HD1 is a nuclear protein. When expressed in yeast, murine HD1 was also detected in the nucleus, although it failed to complement the rpd3⌬ deletion phenotype. HD1 mRNA expression was low in G 0 mouse cells but increased when the cells crossed the G 1 /S boundary after growth stimulation. Immunoprecipitation experiments and functional in vitro assays showed that HD1 protein is associated with histone deacetylase activity. Both HD1 protein levels and total histone deacetylase activity increased upon interleukin-2 stimulation of resting B6.1 cells. When coexpressed with a luciferase reporter construct, HD1 acted as a negative regulator of the Rous sarcoma virus enhancer/promoter. HD1 overexpression in stably transfected Swiss 3T3 cells caused a severe delay during the G 2 /M phases of the cell cycle. Our results indicate that balanced histone acetylation/deacetylation is crucial for normal cell cycle progression of mammalian cells.

Expression deconvolution: A reinterpretation of DNA microarray data reveals dynamic changes in cell populations

“…Deconvolution reveals that cln3⌬ cells show roughly equal proportions of G 1 and S phase cells, but no other phases. For each of the other strains, the observed deconvolution results match the defects observed by other techniques (17)(18)(19)(20)(21), such as the known post-S phase delay of cells overexpressing calmodulin [CMD1 (Tet)], which are defective in nuclear division (22), exhibiting a higher proportion of M and M͞G 1 cells in our analysis.…”

Transcriptional Regulation by the Repressor of Estrogen Receptor Activity via Recruitment of Histone Deacetylases

“…The enzyme is not only essential for mouse development but also for normal proliferation of mouse embryos and embryonic stem cells (23). HDAC1 expression has to be tightly regulated since both overexpression and loss of HDAC1 result in severe disturbance of proliferation and cell cycle progression (23,24). Transcription of the murine HDAC1 gene is induced by cooperative phosphorylation and acetylation signals, allowing both growth factor-dependent activation and feedback regulation (25,26).…”

Modulation of p120E4F transcriptional activity by the Gam1 adenoviral early protein

“…Lanes 1, 3, 6 and 9: 5 mg p120 E4F plus 5 mg myc-HDAC1 plus 5 mg of myc-Gam1; lane 2: 5 mg of myc-Gam1 plus 10 mg of empty vector; lane 4: 5 mg myc-HDAC1 plus 10 mg of empty vector; lane 5: 5 mg myc-HDAC1 plus 5 mg of myc-Gam1 plus 5 mg of empty vector; lane 7: 5 mg p120 E4F plus 10 mg of empty vector; lane 8: 5 mg p120 E4F plus 5 mg myc-HDAC1 plus 5 mg empty vector. Cells were ( 35 S) Met labeled (Amersham) and cellular lysates were immunoprecipitated with anti-LexA antibody (Santa-Cruz) (lane 1), anti-myc antibody (lanes 2 and 3), anti-HDAC1 antibody (Bartl et al, 1997) (lanes 4-6) and anti-HA (lanes 7-9). (b) U2OS cells (1 Â 10 6 ) were transfected with 5 mg of HA-p120 E4F , 5 mg of myc-HDAC1 and 5 mg of myc-Gam1.…”

“…As shown in Figure 2a, in vitro translated p120 E4F bound to GST- HDAC1. To assess whether the two proteins would also interact in vivo, myc-HDAC1 (Bartl et al, 1997) and HA-p120 E4F (Sandy et al, 2000) were expressed in HeLa cells. Immunoprecipitation with anti-myc followed by immunoblotting with anti-HA demonstrated that HDAC1 and p120 E4F interact (Figure 2b, lane 3).…”