The Journal of Cell Biology, Vol 109, 2395-2403, Copyright © 1989 by The Rockefeller University Press

ARTICLES

Differential inhibition of nerve growth factor responses by purine analogues: correlation with inhibition of a nerve growth factor- activated protein kinase

C Volonte, A Rukenstein, DM Loeb and LA Greene
Department of Pathology, College of Physicians and Surgeons of Columbia University, New York 10032.

Purine analogues were used in this study to dissect specific steps in the mechanism of action of nerve growth factor (NGF). Protein kinase N (PKN) is an NGF-activated serine protein kinase that is active in the presence of Mn++. The activity of PKN was inhibited in vitro by purine analogues, the most effective of which was 6-thioguanine (apparent Ki = 6 microM). Several different criteria indicated that 6-thioguanine is not a general inhibitor of protein kinases and that it is relatively specific for PKN. For instance, it did not affect protein kinases A or C and was without effect on the overall level and pattern of protein phosphorylation by either intact or broken PC12 cells. Since purine analogues rapidly and effectively enter cells, they were also assessed for their actions on both transcription-dependent and -independent responses of PC12 cells to NGF. NGF-promoted neurite regeneration was reversibly suppressed by the analogues and at concentrations very similar to those that inhibit PKN. Comparable concentrations of the analogues also blocked NGF-stimulated induction of ornithine decarboxylase activity. In contrast to its inhibition of neurite regeneration and ornithine decarboxylase induction, 6-thioguanine did not suppress NGF-dependent induction of c-fos mRNA expression. Thus, purine analogues such as 6-thioguanine appear capable of differentially suppressing some, but not other actions of NGF. These findings suggest the presence of multiple pathways in the NGF mechanism and that these can be dissected with purine analogues. Moreover, these data are compatible with a role for protein kinase N in certain of these pathways.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Irwin, N., Li, Y.-M., O'Toole, J. E., Benowitz, L. I. (2006). Mst3b, a purine-sensitive Ste20-like protein kinase, regulates axon outgrowth. Proc. Natl. Acad. Sci. USA 103: 18320-18325 [Abstract] [Full Text]  
• Li, Y., Irwin, N., Yin, Y., Lanser, M., Benowitz, L. I. (2003). Axon Regeneration in Goldfish and Rat Retinal Ganglion Cells: Differential Responsiveness to Carbohydrates and cAMP. J. Neurosci. 23: 7830-7838 [Abstract] [Full Text]  
• Chen, P., Goldberg, D. E., Kolb, B., Lanser, M., Benowitz, L. I. (2002). Inosine induces axonal rewiring and improves behavioral outcome after stroke. Proc. Natl. Acad. Sci. USA 99: 9031-9036 [Abstract] [Full Text]  
• Petrausch, B., Tabibiazar, R., Roser, T., Jing, Y., Goldman, D., Stuermer, C. A. O., Irwin, N., Benowitz, L. I. (2000). A Purine-Sensitive Pathway Regulates Multiple Genes Involved in Axon Regeneration in Goldfish Retinal Ganglion Cells. J. Neurosci. 20: 8031-8041 [Abstract] [Full Text]  
• Benowitz, L. I., Goldberg, D. E., Madsen, J. R., Soni, D., Irwin, N. (1999). Inosine stimulates extensive axon collateral growth in the rat corticospinal tract after injury. Proc. Natl. Acad. Sci. USA 96: 13486-13490 [Abstract] [Full Text]  
• Miller, C.S., Danaher, R.J., Jacob, R.J. (1998). Molecular Aspects of Herpes Simplex Virus I Latency, Reactivation, and Recurrence. CROBM 9: 541-562 [Abstract] [Full Text]  
• Abe, T., Morgan, D. A., Gutterman, D. D. (1997). Protective Role of Nerve Growth Factor Against Postischemic Dysfunction of Sympathetic Coronary Innervation. Circulation 95: 213-220 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents