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  • Original Paper
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Physical interaction between CDK9 and B-Myb results in suppression of B-Myb gene autoregulation

Oncogene volume 19, pages 373–379 (2000)Cite this article

Abstract

B-Myb is a transcription factor belonging to the myb family, whose activity has been associated with augmented DNA synthesis and cell cycle progression. We showed recently that B-Myb autoregulates its own expression through promoter transactivation. We report in this study that CDK9, the cyclin T associated kinase, which phosphorylates and activates RNA-Polymerase II, suppresses B-Myb autoregulation through direct interaction with the carboxyl-terminus of the B-Myb protein. Down-regulation of the transactivating ability of B-Myb is independent of the kinase activity of CDK9, because a kinase deficient mutant (dn-CDK9) also represses B-myb gene autoregulation. Overexpression of CDK9 did not result in suppression of p53-dependent transactivation or inhibition of the basal activity of the promoters tested so far, demonstrating that CDK9 is a B-Myb-specific repressor. Rather, transfection of the dominant negative dn-CDK9 construct inhibited the basal activity of the reporter genes, confirming an essential role for CDK9 in gene transcription. In addition, Cyclin T1 restores B-Myb transactivating activity when co-transfected along with CDK9, suggesting that the down-regulatory effect observed on B-Myb is specifically due to CDK9 alone. Thus, our data suggest that CDK9 is involved in the negative regulation of activated transcription mediated by certain transcription factors, such as B-Myb. This may indicate the existence of a feedback loop, mediated by the different activities of CDK9, which links basal with activated transcription.

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Abbreviations

NaCl:

Sodium Chloride

MgCl2:

Magnesium Chloride

HEPES:

N-[2-Hydroxylethyl]-piperazine-N’-[2-ethansulfonic acid]

EGTA:

Ethylenglycol-bis(b-amino-ethylen)N,Nrsquo;-Tetraacetic acid

EDTA:

Ethylenediaminetetraacetic acid

NP-40:

Nonyl Phenoxy Polyetoxy Ethanol

CAPS:

3-[Cycloheximylamino]-1-propanesulfonic acid

PBS:

Phosphate Buffer Saline

TBST:

Tris Buffer Saline+Triton X-100

BSA:

Bovine Serum Albumin

DTT:

Dithyothreitol

NRS:

Normal Rabbit Serum

References

  • Bagella L, MacLachlan TK, Buono RJ, Pisano MM, Giordano A and De Luca A. . 1998 J. Cell. Physiol. 177: 206–213.

  • Bies J, Hoffman B, Amanullah A, Giese T and Wolff L. . 1996 Oncogene 12: 355–363.

  • Bouwmeester T, Guehmann S, el-Baradi T, Kalkbrenner F, van Wijk I, Moelling K and Pieler T. . 1992 Mech. Dev. 37: 57–68.

  • Dahmus ME. . 1996 J. Biol. Chem. 217: 19009–19012.

  • De Falco G and Giordano A. . 1998 J. Cell. Physiol. 177: 501–506.

  • De Luca A, Esposito V, Baldi A, Claudio PP, Fu Y, Caputi M, Pisano M, Baldi F and Giordano A. . 1997 J. Cell. Physiol. 172: 265–273.

  • Gonda TJ and Bishop JM. . 1983 J. Virol. 46: 212–220.

  • Gonda TJ and Metcalf D. . 1984 Nature 310: 249–251.

  • Graña X, De Luca A, Sang N, Fu Y, Claudio PP, Rosenblatt J, Morgan DO and Giordano A. . 1994 Proc. Natl. Acad. Sci. USA 91: 3834–3838.

  • Ho SN, Hunt HD, Orton RM, Pullen JK and Pease LR. . 1989 Gene 77: 51–59.

  • Hoffman-Liebermann B and Liebermann DA. . 1991 Oncogene 6: 903–909.

  • Kalkbrenner F, Guehmann S and Moelling K. . 1990 Oncogene 5: 657–661.

  • Katzen L and Bishop JM. . 1996 Proc. Natl. Acad. Sci. USA 93: 13955–13960.

  • Klempnauer KH and Sippel AE. . 1987 EMBO J. 6: 2719–2725.

  • Klempnauer KH, Ramsay G, Bishop JM, Moscovivi MG, McGrath JP and Levinson AD. . 1983 Proc. Natl. Acad. Sci. USA 33: 345–355.

  • Lam EW and Watson RJ. . 1993 EMBO J. 12: 2705.

  • Lane S, Farlie P and Watson R. . 1997 Oncogene 14: 2445–2453.

  • Lew J, Huang QQ, Qi Z, Winfkein RJ, Aebersold R, Hunt T and Wang JH. . 1994 Nature 371: 423–426.

  • Lyon J, Robinson C and Watson R. . 1994 Crit. Rev. Oncog. 5: 373–378.

  • MacLachlan TK, Sang N, Puri PL, De Luca A, Levrero M and Giordano A. . 1998 J. Cell. Biochem. 71: 467–478.

  • Majello B, Napolitano G, Giordano A and Lania L. . 1999 Oncogene 18: 4598–4605.

  • Napolitano G, Licciardo P, Gallo P, Majello B, Giordano A and Lania L. . 1999 AIDS 13: 1453–1459.

  • Nomura N, Takahashi M, Matsui M, Ishii S, Date T, Sasamoto S and Ishizaki R. . 1988 Nucleic Acid. Res. 16: 11075–11089.

  • Peng J, Zhu Y, Milton JT and Price DH. . 1998 Genes Dev. 12: 755–762.

  • Raschella’ G, Negroni A, Sala A, Pucci S, Romeo A and Calabretta B. . 1995 J. Biol. Chem. 270: 8540–8545.

  • Raschella’ G, Tanno B, Bonetto F, Amendola R, Battista T, De Luca A, Giordano A and Paggi MG. . 1997 J. Cell. Biochem. 67: 297–303.

  • Reiss K, Travali S, Calabretta B and Baserga R. . 1991 J. Cell. Physiol. 14: 338–343.

  • Romano G, Kasten M, De Falco G, Micheli P, Khalili K and Giordano A. . 1999 J. Cell. Biochem. 75: 357–368.

  • Sala A and Watson R. . 1999 J. Cell. Physiol. 179: 245–250.

  • Sala A, Saitta P, De Luca A, Casella I, Lewis RE, Watson R and Peschle C. . 1999 Oncogene 18: 1333–1339.

  • Sala A, Casella I, Grasso L, Bellon T, Reed JC, Miyashita T and Peschle C. . 1996a Cancer Res. 56: 1991–1996.

  • Sala A, De Luca A, Giordano A and Peschle C. . 1996b J. Biol. Chem. 271: 28738–28740.

  • Sala A, Kundu M, Casella I, Engelhard A, Calabretta B, Grasso L, Paggi MG, Giordano A, Watson RJ, Khalili K and Peschle C. . 1997 Proc. Natl. Acad. Sci. USA 94: 532–536.

  • Sanger F, Nicklen S and Coulson AR. . 1977 Proc. Natl. Acad. Sci. USA 74: 5463–5467.

  • Saville MK and Watson RJ. . 1988 Adv. Cancer Res. 72: 109–140.

  • Sitzmann J, Noben-Trauth K, Kamano H and Klempnauer KH. . 1996 Oncogene 12: 1889–1894.

  • Tsai LH, Delalle I, Cavines VJ, Chae T and Harlow E. . 1994 Nature 371: 419–423.

  • Watson RJ, Robinson C and Lam EW. . 1993 Nucleic. Acid. Res. 21: 267–272.

  • Wei P, Gerber ME, Fang SM, Fischer WH and Jones K. . 1998 Cell 92: 451–462.

  • Yang X, Gold MO, Tang DN, Lewis DE, Aguilar-Cordova E, Rice AP and Herrman C. . 1997 Proc. Natl. Acad. Sci. USA 94: 12331–12336.

  • Ziebold U, Bartsch O, Marais R, Ferrari S and Klempnauer KH. . 1997 Curr. Biol. 7: 253–260.

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Acknowledgements

This work has been supported in part by grants from the National Institute of Health (A Giordano and B Calabretta). G De Falco was partially supported from an AIDS fellowship from the ‘Istituto Superiore di Sanita’. PP Claudio is the recipient of a fellowship from the ‘Associazione Leonardo di Capua’, Napoli, Italy. A De Luca is the recipient of a grant FIRC. L Bagella is supported by a Dottorato di Ricerca in Biologia Diagnostica Quantitiva from the Universita di Siena.

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Author notes

  1. Giulia De Falco and Luigi Bagella: G De Falco and L Bagella contributed equally

Authors and Affiliations

  1. Department of Pathology, Anatomy & Cell Biology, Jefferson Medical College of Thomas Jefferson University, Philadelphia, 19107, Pennsylvania, PA, USA

    Giulia De Falco, Luigi Bagella, Pier Paolo Claudio, Yan Fu & Antonio Giordano

  2. Department of ‘Scienze Odontostomatologiche e Maxillo Facciali’, Universita’ di Napoli ‘Federico II’, via Pansini 5, Naples, Italy

    Pier Paolo Claudio

  3. Laboratory for Cell Metabolism and Pharmacokinetics, Center for Experimental Research, Regina Elena Institute, Via delle Messi d’Oro 156, Rome, 00158, Italy

    Antonio De Luca

  4. Department of Microbiology, Kimmel Cancer Institute, Jefferson Medical College of Thomas Jefferson University, Philadelphia, 19107, Pennsylvania, PA, USA

    Bruno Calabretta

  5. Department of Molecular Pharmacology and Pathology, Consorzio Mario Negri Sud, 66030 S. Mario Imbaro (CH), Italy

    Arturo Sala

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  1. Giulia De Falco
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De Falco, G., Bagella, L., Claudio, P. et al. Physical interaction between CDK9 and B-Myb results in suppression of B-Myb gene autoregulation. Oncogene 19, 373–379 (2000). https://doi.org/10.1038/sj.onc.1203305

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