Autoregulation of Mouse Histone Deacetylase 1 Expression

Division of Molecular Biology, Institute of Medical Biochemistry, University of Vienna, Vienna Biocenter, A-1030 Vienna, Austria¹

^{Corresponding author. Mailing address: Christian Seiser Institute of Medical Biochemistry, Division of Molecular Biology, University of Vienna, Vienna Biocenter, Dr. Bohr-Gasse 9/2, A-1030 Vienna, Austria. Phone: 431 4277 61770. Fax: 431 4277 9617. E-mail:}ta.ca.eivinu.lom@sc.

Received 2003 May 28; Accepted 2003 Jul 2.

Abstract

Histone deacetylase 1 (HDAC1) is a major regulator of chromatin structure and gene expression. Tight control of HDAC1 expression is essential for development and normal cell cycle progression. In this report, we analyzed the regulation of the mouse HDAC1 gene by deacetylases and acetyltransferases. The murine HDAC1 promoter lacks a TATA box consensus sequence but contains several putative SP1 binding sites and a CCAAT box, which is recognized by the transcription factor NF-Y. HDAC1 promoter-reporter studies revealed that the distal SP1 site and the CCAAT box are crucial for HDAC1 promoter activity and act synergistically to constitute HDAC1 promoter activity. Furthermore, these sites are essential for activation of the HDAC1 promoter by the deacetylase inhibitor trichostatin A (TSA). Chromatin immunoprecipitation assays showed that HDAC1 is recruited to the promoter by SP1 and NF-Y, thereby regulating its own expression. Coexpression of acetyltransferases elevates HDAC1 promoter activity when the SP1 site and the CCAAT box are intact. Increased histone acetylation at the HDAC1 promoter region in response to TSA treatment is dependent on binding sites for SP1 and NF-Y. Taken together, our results demonstrate for the first time the autoregulation of a histone-modifying enzyme in mammalian cells.

Abstract

In eukaryotic cells, DNA is complexed with core histones and other proteins in the form of chromatin. The basic repeating unit of chromatin, the nucleosome, is built of two copies of each of the four core histones, H2A, H2B, H3, and H4, wrapped by 146 bp of DNA. This organization allows the efficient packaging of genomic DNA into the nucleus but also has a negative impact on gene expression. To overcome this nucleosomal repression, the N-terminal tails of core histones are targets for multiple modifications, such as acetylation, phosphorylation, and methylation, which can modulate chromatin compaction. The best-studied modification of core histones is the reversible acetylation of conserved lysine residues within the N termini. Acetylation results in reduced interaction between positively charged histone tails and negatively charged DNA. Histone deacetylation is believed to result in chromatin condensation, whereas acetylation correlates with increased accessibility to genes for the transcription machinery.

Two types of enzymes, the histone acetyltransferases (HATs) and the histone deacetylases (HDACs), control the acetylation of histones and other proteins. More than a dozen mammalian histone deacetylases have been identified in recent years, and they have been classified into three groups according to their homology with the yeast enzymes Rpd3, Hda1, and Sir2 (11, 19). Class I enzymes seem to be involved in more general cellular processes, whereas class II enzymes might have more tissue-specific functions. The third mammalian HDAC class is made up of enzymes with homology to the NAD-dependent deacetylase Sir2. Mammalian Sir2 was recently shown to deacetylate p53, thereby controlling stress response and cell survival (21, 24, 38).

The class I enzyme HDAC1 was the first mammalian deacetylase identified (37). Numerous transcription factors, including regulators of the cell cycle, differentiation, and development, have been shown to associate with HDAC1, thereby mediating the repression of specific target genes (1, 7, 27). Previous work from our laboratory indicated a role of mouse HDAC1 in the regulation of proliferation and development. For instance, it has been shown that the expression of HDAC1 is induced upon growth factor activation of mouse T cells and fibroblasts (3, 13). In addition, HDAC1 levels were found to be elevated in highly proliferative tissues, embryonic stem cells, and several transformed cell lines (3, 20), suggesting a link between HDAC1 function and proliferation. In accordance with this idea, disruption of the HDAC1 gene resulted in reduced proliferation of mouse embryos and embryonic stem cells (20), whereas overexpression of HDAC1 led to impaired proliferation of murine fibroblasts (3). Taken together, these results indicate that a tightly controlled cell-type-specific expression of HDAC1 is crucial for unrestricted proliferation.

Recent findings point to the existence of a control system that modulates cellular deacetylase activities via a regulatory feedback mechanism. The expression of HDAC1 and certain other mammalian histone deacetylases is increased in response to deacetylase inhibitor treatment (10, 39). Furthermore, the HDAC1 gene was recently shown to be activated by the cooperation of acetylating and phosphorylating signals, resulting in phosphoacetylation of HDAC1 promoter-associated histone H3 (13). These data demonstrated that the chromatin modifier HDAC1 is regulated by mechanisms involving changes in chromatin structure.

Here, we directly evaluate the roles of acetylases and deacetylases in the regulation of HDAC1 promoter activity. We show that binding sites important for HDAC1 promoter activity are also essential for activation of the promoter by the deacetylase inhibitor trichostatin A (TSA). Further, we demonstrate that SP1 and NF-Y transcription factors can bind to these consensus sequences and can recruit HDAC1 to its own promoter. The HDAC1-mediated repression is counteracted by histone acetyltransferases, such as p300 and P/CAF. This feedback loop provides a perfect mechanism for the precise regulation of HDAC1 levels in mammalian cells.

Acknowledgments

We thank G. Zupkovitz for cultivating the ES cells, G. Weitzer for the GenomeWalker kit, G. Suske and R. Mantovani for antibodies, T. F. Osborne for the Drosophila expression constructs, E. Wintersberger for the NF-Y constructs and antibodies, H. Rotheneder for antibodies, T. Sauer for fluorescence-activated cell sorter analyses, and E. Wintersberger, H. Rotheneder, and R. Moriggl for helpful comments on the manuscript.

This work was supported by the Austrian FWF (grant P14909-GEN) and the Herzfelder Family Foundation.

Acknowledgments

REFERENCES

References

1. Ahringer, J. 2000. NuRD and SIN3 histone deacetylase complexes in development. Trends Genet.16:351-356. [[PubMed]
2. Baker, A., M. Saltik, H. Lehrmann, I. Killisch, V. Mautner, G. Lamm, G. Christofori, and M. Cotten. 1997. Polyethylenimine (PEI) is a simple, inexpensive and effective reagent for condensing and linking plasmid DNA to adenovirus for gene delivery. Gene Ther.4:773-782. [[PubMed]
3. Bartl, S., J. Taplick, G. Lagger, H. Khier, K. Kuchler, and C. Seiser. 1997. Identification of mouse histone deacetylase 1 as a growth factor-inducible gene. Mol. Cell. Biol.17:5033-5043.
4. Billon, N., D. Carlisi, M. B. Datto, L. A. van Grunsven, A. Watt, X. F. Wang, and B. B. Rudkin. 1999. Cooperation of Sp1 and p300 in the induction of the CDK inhibitor p21WAF1/CIP1 during NGF-mediated neuronal differentiation. Oncogene18:2872-2882. [[PubMed]
5. Braun, H., R. Koop, A. Ertmer, S. Nacht, and G. Suske. 2001. Transcription factor Sp3 is regulated by acetylation. Nucleic Acids Res.29:4994-5000.
6. Choi, H., J. Lee, J. Park, and Y. Lee. 2002. Trichostatin A, a histone deacetylase inhibitor, activates the IGFBP-3 promoter by upregulating Sp1 activity in hepatoma cells: alteration of the Sp1/Sp3/HDAC1 multiprotein complex. Biochem. Biophys. Res. Commun.296:1005-1012. [[PubMed]
7. Cress, W. D., and E. Seto. 2000. Histone deacetylases, transcriptional control, and cancer. J. Cell. Physiol.184:1-16. [[PubMed]
8. Dangond, F., and S. R. Gullans. 1998. Differential expression of human histone deacetylase mRNAs in response to immune cell apoptosis induction by trichostatin A and butyrate. Biochem. Biophys. Res. Commun.247:833-837. [[PubMed]
9. Doetzlhofer, A., H. Rotheneder, G. Lagger, M. Koranda, V. Kurtev, G. Brosch, E. Wintersberger, and C. Seiser. 1999. Histone deacetylase 1 can repress transcription by binding to Sp1. Mol. Cell. Biol.19:5504-5511.
10. Gray, S. G., and T. J. Ekstrom. 1998. Effects of cell density and trichostatin A on the expression of HDAC1 and p57Kip2 in Hep 3B cells. Biochem. Biophys. Res. Commun.245:423-427. [[PubMed]
11. Gray, S. G., and T. J. Ekström. 2001. The human histone deacetylase family. Exp. Cell Res.262:75-83. [[PubMed]
12. Hagen, G., S. Muller, M. Beato, and G. Suske. 1994. Sp1-mediated transcriptional activation is repressed by Sp3. EMBO J.13:3843-3851.
13. Hauser, C., B. Schuettengruber, S. Bartl, G. Lagger, and C. Seiser. 2002. Activation of the HDAC1 gene by cooperative histone phosphorylation and acetylation. Mol. Cell. Biol.22:7820-7830.
14. Hu, Z., S. Jin, and K. W. Scotto. 2000. Transcriptional activation of the MDR1 gene by UV irradiation. Role of NF-Y and Sp1. J. Biol. Chem.275:2979-2985. [[PubMed]
15. Inoue, T., J. Kamiyama, and T. Sakai. 1999. Sp1 and NF-Y synergistically mediate the effect of vitamin D(3) in the p27(Kip1) gene promoter that lacks vitamin D response elements. J. Biol. Chem.274:32309-32317. [[PubMed]
16. Jin, S., and K. W. Scotto. 1998. Transcriptional regulation of the MDR1 gene by histone acetyltransferase and deacetylase is mediated by NF-Y. Mol. Cell. Biol.18:4377-4384.
17. Karlseder, J., H. Rotheneder, and E. Wintersberger. 1996. Interaction of Sp1 with the growth- and cell cycle-regulated transcription factor E2F. Mol. Cell. Biol.16:1659-1667.
18. Khier, H., S. Bartl, B. Schuettengruber, and C. Seiser. 1999. Cloning and characterization of the mouse histone deacetylase 1 gene: integration of a retrovirus in 129SV mice. Biochim. Biophys. Acta1489:365-373. [[PubMed]
19. Khochbin, S., and H. Y. Kao. 2001. Histone deacetylase complexes: functional entities or molecular reservoirs. FEBS Lett.494:141-144. [[PubMed]
20. Lagger, G., D. O'Carroll, M. Rembold, H. Khier, J. Tischler, G. Weitzer, B. Schuettengruber, C. Hauser, R. Brunmeir, T. Jenuwein, and C. Seiser. 2002. Essential function of histone deacetylase 1 in proliferation control and CDK inhibitor repression. EMBO J.21:2672-2681.
21. Langley, E., M. Pearson, M. Faretta, U. M. Bauer, R. A. Frye, S. Minucci, P. G. Pelicci, and T. Kouzarides. 2002. Human SIR2 deacetylates p53 and antagonizes PML/p53-induced cellular senescence. EMBO J.21:2383-2396.
22. Li, Q., M. Herrler, N. Landsberger, N. Kaludov, V. V. Ogryzko, Y. Nakatani, and A. P. Wolffe. 1998. Xenopus NF-Y pre-sets chromatin to potentiate p300 and acetylation-responsive transcription from the Xenopus hsp70 promoter in vivo. EMBO J.17:6300-6315.
23. Liang, F., F. Schaufele, and D. G. Gardner. 2001. Functional interaction of NF-Y and Sp1 is required for type a natriuretic peptide receptor gene transcription. J. Biol. Chem.276:1516-1522. [[PubMed]
24. Luo, J., A. Y. Nikolaev, S.-I. Imai, A. Shiloh, L. Guarente, and W. Gu. 2001. Negative control of p53 by Sir2α promotes cell survival under stress. Cell107:137-148. [[PubMed]
25. Maity, S. N., and B. de Crombrugghe. 1998. Role of the CCAAT-binding protein CBF/NF-Y in transcription. Trends Biochem. Sci.23:174-178. [[PubMed]
26. Manni, I., G. Mazzaro, A. Gurtner, R. Mantovani, U. Haugwitz, K. Krause, K. Engeland, A. Sacchi, S. Soddu, and G. Piaggio. 2001. NF-Y mediates the transcriptional inhibition of the cyclin B1, cyclin B2, and cdc25C promoters upon induced G2 arrest. J. Biol. Chem.276:5570-5576. [[PubMed]
27. Ng, H. H., and A. Bird. 2000. Histone deacetylases: silencers for hire. Trends Biochem. Sci.25:121-126. [[PubMed]
28. Pan, J., and R. P. McEver. 1993. Characterization of the promoter for the human P-selectin gene. J. Biol. Chem.268:22600-22608. [[PubMed]
29. Peng, Y., and NJahroudi. 2003. The NFY transcription factor inhibits von Willebrand factor promoter activation in non-endothelial cells through recruitment of histone deacetylases. J. Biol. Chem.278:8385-8394. [[PubMed][Google Scholar]
30. Roder, K., S. S. Wolf, K. F. Beck, and M. Schweizer. 1997. Cooperative binding of NF-Y and Sp1 at the DNase I-hypersensitive site, fatty acid synthase insulin-responsive element 1, located at −500 in the rat fatty acid synthase promoter. J. Biol. Chem.272:21616-21624. [[PubMed]
31. Roder, K., S. S. Wolf, K. J. Larkin, and M. Schweizer. 1999. Interaction between the two ubiquitously expressed transcription factors NF-Y and Sp1. Gene234:61-69. [[PubMed]
32. Salsi, V., G. Caretti, M. Wasner, W. Reinhard, U. Haugwitz, K. Engeland, and R. Mantovani. 2003. Interactions between p300 and multiple NF-Y trimers govern cyclin B2 promoter function. J. Biol. Chem.278:6642-6650. [[PubMed]
33. Sorensen, P., and EWintersberger. 1999. Sp1 and NF-Y are necessary and sufficient for growth-dependent regulation of the hamster thymidine kinase promoter. J. Biol. Chem.274:30943-30949. [[PubMed][Google Scholar]
34. Sowa, Y., T. Orita, S. Minamikawa, K. Nakano, T. Mizuno, H. Nomura, and T. Sakai. 1997. Histone deacetylase inhibitor activates the WAF1/Cip1 gene promoter through the Sp1 sites. Biochem. Biophys. Res. Commun.241:142-150. [[PubMed]
35. Sun, J. M., H. Y. Chen, M. Moniwa, D. W. Litchfield, E. Seto, and J. R. Davie. 2002. The transcriptional repressor Sp3 is associated with CK2 phosphorylated histone deacetylase 2. J. Biol. Chem.277:35783-35786. [[PubMed]
36. Suske, G. 1999. The Sp-family of transcription factors. Gene238:291-300. [[PubMed]
37. Taunton, J., C. A. Hassig, and S. L. Schreiber. 1996. A mammalian histone deacetylase related to the yeast transcriptional regulator Rpd3p. Science272:408-411. [[PubMed]
38. Vaziri, H., S. K. Dessain, E. Ng Eaton, S. I. Imai, R. A. Frye, T. K. Pandita, L. Guarente, and R. A. Weinberg. 2001. hSIR2(SIRT1) functions as an NAD-dependent p53 deacetylase. Cell107:149-159. [[PubMed]
39. Verdel, A., and SKhochbin. 1999. Identification of a new family of higher eukaryotic histone deacetylases—coordinate expression of differentiation-dependent chromatin modifiers. J. Biol. Chem.274:2440-2445. [[PubMed][Google Scholar]
40. Xu, Y., D. Banville, H. F. Zhao, X. Zhao, and S. H. Shen. 2001. Transcriptional activity of the SHP-1 gene in MCF7 cells is differentially regulated by binding of NF-Y factor to two distinct CCAAT-elements. Gene269:141-153. [[PubMed]
41. Yamada, K., T. Tanaka, K. Miyamoto, and T. Noguchi. 2000. Sp family members and nuclear factor-Y cooperatively stimulate transcription from the rat pyruvate kinase M gene distal promoter region via their direct interactions. J. Biol. Chem.275:18129-18137. [[PubMed]
42. Yun, J., H. D. Chae, H. E. Choy, J. Chung, H. S. Yoo, M. H. Han, and D. Y. Shin. 1999. p53 negatively regulates cdc2 transcription via the CCAAT-binding NF-Y transcription factor. J. Biol. Chem.274:29677-29682. [[PubMed]
43. Zeng, Y. Y., C. M. Tang, Y. L. Yao, W. M. Yang, and E. Seto. 1998. Cloning and characterization of the mouse histone deacetylase-2 gene. J. Biol. Chem.273:28921-28930. [[PubMed]
44. Zhang, Y., and M. L. Dufau. 2002. Silencing of transcription of the human luteinizing hormone receptor gene by histone deacetylase-mSin3A complex. J. Biol. Chem.277:33431-33438. [[PubMed]
45. Zhao, S., K. Venkatasubbarao, S. Li, and J. W. Freeman. 2003. Requirement of a specific Sp1 site for histone deacetylase-mediated repression of transforming growth factor beta type II receptor expression in human pancreatic cancer cells. Cancer Res.63:2624-2630. [[PubMed]