Abstract
Several types of cellular proteins can be modified by farnesylation and nitrosylation, of which the most significant is Ras. We used manumycin, a farnesyltransferase inhibitor, and L-NAME (Nω-nitro-l-arginine methyl ester), a nitric oxide synthase (NOS) inhibitor, for characterization of Ras-dependent downstream targets activities. Our results suggest that change of the steady-state levels of nitric oxide and inhibition of farnesylation modified the activities of several transcription factors. We have found that the inhibition of farnesylation by manumycin decreased the DNA-binding activity of nuclear factor (NF)-κB, did not change the DNA-binding activities of STAT, Sp1, ATF-2, and CREB, and increased the activities of c-Fos, JunD, and c-Jun. Under such conditions, phosphorylation of Akt was decreased, whereas phosphorylation of extracellular signal-regulated kinase (ERK) was increased and phosphorylation of JNK did not change. Furthermore, our results show that reduction of intracellular concentration of nitric oxides by L-NAME increases the activities of c-Fos, ATF-2 and JunD and decreases the activities of CREB, STAT, Sp1, and c-Jun. The activities of all of these transcription factors are restored to normal levels in the presence of manumycin, suggesting that simultaneous modifications of proteins by farnesylation and nitrosylation change the direction of Ras-controlled downstream pathways. Our results provide further evidence of the significance of posttranslational modifications of Ras for the specificity of transducing cascade networks and physiological outcome.
Similar content being viewed by others
References
Baker T. A., Booden M. A., and Buss J. E. (2000) S-Nitrosocysteine increases palmitate turnover on Ha-Ras in NIH 3T3 cells. J. Biol. Chem. 275, 22,037–22,047.
Booden M., Sakaguchi D. S., and Buss J. E. (2000) Mutation of Ha-Ras C terminus changes effector pathway utilization. J. Biol. Chem. 275, 23,559–23,568.
Caccavelli L., Manfroid I., Martial J. A., and Muller M. (1998) Transcription factor AP1 is involved in basal and okadaic acid-stimulated activity of the human PRL promoter. Mol. Endocrinol. 12(8), 1215–1227.
Celec P. (2004) Nuclear factor kappa B—molecular biomedicine:the next generation. Biomed. Pharmacother. 58, 365–371.
Darnell J. E. Jr. (1997) STATs and gene regulation. Science 277, 1630–1635.
Garrington T. P. and Johnson G. L. Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr. Opin. Cell Biol. 11(1999), 211–218.
Gupta S., Campbell D., Derijard B., and Davis R. J. (1995) Transcription factor ATF2 regulation by the JNK signal transduction pathway. Science 267, 389–393.
Jiang K., Coppola D., Crespo N. C., et al. (2000) The phosphoinositide 3-OH kinase/AKT2 pathway as a critical target for farnesyltransferase inhibitor-induced apoptosis. Mol. Cell. Biol. 20, 139–148.
Kim O. S., Park E. J., Joe E-H., and Jou I. (2002) JAK-STAT Signaling mediates gangliosides-induced inflammatory responses in brain microglial cells. J. Biol. Chem. 277, 40,594–40,601.
Lander H. M., Milbank A. J., Tauras J. M., et al. (1996) Redox regulation of cell signalling. Nature (London) 381, 380–381.
Leppä S., Eriksson M., Saffrich R., Ansorge W., and Bohmann D. (2001) Complex functions of AP-1 transcription factors in differentiation and survival of PC12 cells. Mol. Cell. Biol. 21, 4369–4378.
Lilienbaum A. and Israel A. (2003) From calcium to NF-kB signaling pathways in neurons. Mol. Cell. Biol. 23(8), 2680–2698.
Mallis R. J., Buss J. E., and Thomasi J. A. (2001) Oxidative modification of H-ras: S-thiolation and S-nitrosylation of reactive cysteines. Biochem. J. 355, 145–153.
Mao X., Moerman A. M., and Barger S. W. (2002) Neuronal kB-binding factors consist of Sp1-related proteins. Functional implications for autoregulation of N-methyl-D-aspartate receptor-1 expression. J. Biol. Chem. 277, 44,911–44,919.
Matthews J. R., Botting C. H., Panico M., Morris H. R., and Hay R. T. (1996) Inhibition of NF-B DNA binding by nitric oxide. Nucleic Acids Res. 24, 2236–2242.
Melnikoval N. and Gardner P. D. (2001) The signal transduction pathway underlying ion channel gene regulation by Sp1-c-Jun interactions. J. Biol. Chem. 276, 19,040–19,045.
Miyazaki T., Takaoka A., Nogueira L., et al. (1998) Pyk2 is a downstream mediator of the IL-2 receptor coupled Jak signaling pathway. Genes Dev. 12, 770–775.
Mott H. R., Carpenter J. W., and Campbell S. L. (1997) Structural and functional analysis of a mutant Ras protein that is insensitive to nitric oxide activation. Biochemistry 36, 3640–3644.
Olson M. F. and Marais R. (2000) Ras protein signalling. Semin. Immunol. 12, 63–73.
Raines K. W., Cao G-L., Porsuphatana S., Tsai P., Rosen G. M., and Shapiro P. (2004) Nitric oxide inhibition of ERK1/2 activity in cells expressing neuronal nitricoxide synthase. J. Biol. Chem. 279(6), 3933–3940.
Ryu H., Lee J., Zaman K., et al. (2003) Sp1 and Sp3 are oxidative stress-inducible, antideath transcription factors in cortical neurons. J. Neurosci. 23, 3597–3606.
So H. S., Park R. K., Kim M. S., et al. (1998) Nitric oxide inhibits c-Jun N-terminal kinase 2 (JNK2) via S-nitrosylation. Biochem. Biophys. Res. Commun. 247, 809–813.
Tabuchi A., Sano K., Oh E., Tsuchiya T., and Tsuda M. (1994) Modulation of AP-1 activity by nitric oxide (NO) in vitro: NO-mediated modulation of AP-1. FEBS Lett. 351, 123–127.
Williams J. G., Pappu K., and Campbell S. L. (2003) Structural and biochemical studies of p21Ras S-nitrosylation and nitric oxide-mediated guanine nucleotide exchange. Proc. Natl. Acad. Sci. U. S. A. 100, 6376–6381.
Wolfman J. C., Palmby T., Der C. J., and Wolfman A. (2002) Cellular N-Ras promotes cell survival by down-regulation of Jun N-terminal protein kinase and p38. Mol. Cell. Biol. 22(5), 1589–1606.
Yamagishi S., Yamada M., Koshimizu H., et al. (2003) Apoptosis-signal regulating kinase-1 is involved in the low potassium-induced activation of p38 mitogen-activated protein kinase and c-Jun in cultured cerebellar granule neurons. J. Biochem. 133, 719–724.
Zeng O., Si X., Horstmann H., Xu Y., Hong W., and Pallen C. J. (2000) Prenylation-dependent Association of Protein-tyrosine Phosphatases PRL-1, -2, and -3 with the plasma membrane and the early endosome. J. Biol. Chem. 275(28), 21,444–21,452.
Zhang J., Wang S., Wesley R. A., and Danner R. L. (2003) Adjacent sequence controls the response polarity of nitric oxide-sensitive Sp factor binding sites. J. Biol. Chem. 278, 29,192–29,200.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhuravliova, E., Barbakadze, T., Narmania, N. et al. Inhibition of nitric oxide synthase and farnesyltransferase change the activities of several transcription factors. J Mol Neurosci 31, 281–287 (2007). https://doi.org/10.1385/JMN:31:03:281
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/JMN:31:03:281