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Functional inactivation of Cdk9 through oligomerization chain reaction

Oncogene volume 22, pages 4882–4888 (2003)Cite this article

Abstract

The oligomerization chain reaction (OCR) strategy is a recently described technique for inactivation of target proteins that function as homoassociate complexes. This novel strategy is based on the fusion of self-associating coiled-coil (CC) domain of the nuclear factor promyelocytic leukemia (PML) to target proteins. Here, we present the successful application of the OCR strategy for inactivation of the heterodimeric Cdk9/cyclin T1 complex. Cyclin T1/Cdk9 (P-TEFb) complex is a positive regulator of gene transcription, whose function is underlined by the ability to phosphorylate the carboxyl-terminal domain (CTD) of the RNA polymerase II conferring productive transcript elongation. Fusion of the CC domain to Cdk9 leads to the formation of high molecular complexes to which the endogenous cyclin T1 is recruited. The CC-Cdk9 chimera effectively inhibits HIV-1 Tat activation, whose transcription activity is exquisitely dependent upon cyclin T1/Cdk9 function. Furthermore, expression of CC-Cdk9 protein inhibits cell proliferation, as shown by colony-formation assay. Collectively, our findings add further support to the OCR strategy for functional inactivation of hetero-associated factors such as the Cdk9/cyclin T1 complex, and highlight a putative function of Cdk9 in cell growth control.

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Acknowledgements

We thank Saverio Minucci and Pier Giuseppe Pelicci for reagents and helpful discussions. This work was supported by grants from the Italian Association for Cancer Research (AIRC), the I.S.S. (Programma Nazionale di Ricerca AIDS, Grant 40B.53) to LL.

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Authors and Affiliations

  1. Department of Genetics, Molecular and General Biology, University of Naples ‘Federico II’, Naples, Italy

    Giuliana Napolitano, Alberto Mazzocco, Alessandro Fraldi, Barbara Majello & Luigi Lania

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  1. Giuliana Napolitano
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  2. Alberto Mazzocco
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  3. Alessandro Fraldi
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  4. Barbara Majello
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  5. Luigi Lania
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Correspondence to Luigi Lania.

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Napolitano, G., Mazzocco, A., Fraldi, A. et al. Functional inactivation of Cdk9 through oligomerization chain reaction. Oncogene 22, 4882–4888 (2003). https://doi.org/10.1038/sj.onc.1206785

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