Regulation of Human mRNA Transcription by NFAT and AP-1 Proteins:
The mammalian immune system represents a unique model for studying the importance of transcriptional regulation in governing cell growth and differentiation. Interleukin-2 is a cytokine that acts as an autocrine growth factor promoting the proliferation and development of T cells during the immune response to bacterial and viral infection, as well as tumorigenesis. The IL-2 promoter is relatively compact for mammalian genes, since proper regulation of IL-2 transcription requires only 340 bp of DNA surrounding the transcription start site. We have investigated the roles of both cis-regulatory elements and trans-regulatory factors (e.g., NFAT, AP1) in transcription at the IL-2 promoter using in vitro transcription experiments and assays in T cells. We have found that NFAT1 and cJun homodimers have a unique ability to interact with one another and that this interaction is critical for the synergistic activation of IL-2 transcription. We are now studying the role of these transcription factors in mediating synergy at other promoters and in other biological systems.
Walters, R.D., Drullinger, L.F., Kugel, J.F., Goodrich, J.A. (2013). NFATc2 recruits cJun homodimers to an NFAT site to synergistically activate interleukin-2 transcription. Mol Immunol. In press.
Nguyen, T.N., Kim, L.J., Walters, R.D., Drullinger, L.F., Lively T.N., Kugel, J.F., and Goodrich, J.A. (2010) The C-terminal region of human NFATc2 binds cJun to synergistically activate interleukin-2 transcription. Mol Immunol. 47(14):2314-22
Weaver, J.R., Good, K., Walters, R.D., Kugel, J.F., and Goodrich, J.A. (2007). Characterization of the sequence and architectural constraints of the regulatory and core regions of the human interleukin-2 promoter. Mol. Immunol. 44: 2813-2819.
Hieb, A.R., Baran, S., Goodrich, J.A., and Kugel, J.F. (2006). An 8 nt RNA triggers a rate-limiting shift of RNA polymerase II complexes into elongation. EMBO J. 25: 3100-3109.
Nguyen, T.N. and Goodrich, J.A. (2006). Immobilized protein-protein interaction assays: Eliminating false positive interactions caused by contaminating nucleic acid. Nature Methods. 3: 135-139.
Weaver, J.R., Kugel, J.F., and Goodrich, J.A. (2005). The sequence at specific positions in the early transcribed region sets the rate of transcript synthesis by RNA polymerase II in vitro. J. Biol. Chem. 280: 39860-39869.
Lively, T.N., Nguyen, T.N., Galasinski, S.K., and Goodrich, J.A. (2004). The basic leucine zipper domain of cJun functions in transcriptional activation through interaction with the N terminus of hsTAF1 (human TAFII250). J. Biol. Chem. 279: 26257-26265.
Ferguson, H.A., Kugel, J.F., and Goodrich, J.A. (2001). Kinetic and mechanistic analysis of the RNA polymerase II transcription reaction at the human interleukin-2 promoter. J. Mol. Biol. 314: 993-1006.
Ferguson, H.A. and Goodrich, J.A. (2001). Expression and purification of recombinant human c Fos/c-Jun that is highly active in DNA binding and transcriptional activation in vitro. Nucleic Acids Res. 29: E98 (6 pages).
Lively, T.N., Ferguson, H.A., Galasinski, S.K., Seto, A.G., and Goodrich, J.A. (2001). c Jun binds the N terminus of human TAFII250 to derepress RNA polymerase II transcription in vitro. J. Biol. Chem. 276: 25582-25588.
Kim, L.J., Seto, A.G., Nguyen, T.N., and Goodrich, J.A. (2001). Human TAFII130 is a coactivator for NFATp. Mol. Cell. Biol. 21: 3503-3513.
Galasinski, S.K., Lively, T.N., Grebe de Barron, A., and Goodrich, J.A. (2000). Acetyl coenzyme A stimulates RNA polymerase II transcription and promoter binding by transcription factor IID in the absence of histones. Mol. Cell. Biol. 20: 1923-1930.