JCB logo
PeproTech: Cell Culture Supplements
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 522K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Vandevoorde, V.
Right arrow Articles by Fiers, W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vandevoorde, V.
Right arrow Articles by Fiers, W.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/1997//1627 $5.00
The Journal of Cell Biology, Volume 137, Number 7, , 1997 1627-1638

Article

Induced Expression of Trimerized Intracellular Domains of the Human Tumor Necrosis Factor (TNF) p55 Receptor Elicits TNF Effects



Veronique Vandevoorde, Guy Haegeman, and Walter Fiers

Laboratory of Molecular Biology, Flanders Interuniversity Institute for Biotechnology and University of Ghent, B-9000 Ghent, Belgium

The various biological activities of tumor necrosis factor (TNF) are mediated by two receptors, one of 55 kD (TNF-R55) and one of 75 kD (TNF-R75). Although the phenotypic and molecular responses elicited by TNF in different cell types are fairly well characterized, the signaling pathways leading to them are so far only partly understood. To further unravel these processes, we focused on TNF-R55, which is responsible for mediating most of the known TNF effects. Since several studies have demonstrated the importance of receptor clustering and consequently of close association of the intracellular domains for signaling, we addressed the question of whether clustering of the intracellular domains of TNF-R55 (TNF-R55i) needs to occur in structural association with the inner side of the cell membrane, where many signaling mediators are known to reside. Therefore, we investigated whether induced intracellular clustering of only TNF-R55i would be sufficient to initiate and generate a full TNF response, without the need for a full-length receptor molecule or a transmembrane region. Our results provide clear evidence that inducible forced trimerization of either TNF-R55i or only the death domain elicits an efficient TNF response, comprising activation of the nuclear factor {kappa}B, induction of interleukin-6, and cell killing.

1. Abbreviations used in this paper: ActD, actinomycin D; CAT, chloramphenicol acetyltransferase; DD, death domain; IFN, interferon; IL-6, interleukin-6; IPTG, isopropyl β-D-thiogalactopyranoside; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NF-{kappa}B, nuclear factor {kappa}B; PI, propidium iodide; PSI, Cbz-Ile-Glu(O-t-Bu)-Ala-leucinal; TNF, tumor necrosis factor; TNF-R55, 55-kD TNF receptor; h- and mTNF-R55, human and murine TNF-R55; TNF-R55i, intracellular domain of TNF-R55; TNF-R75, 75-kD TNF receptor.

We are especially indebted to Dr. C. Weissmann for providing pSP64Mx and an IFN-{alpha} (A/D) preparation. We further thank Dr. G. Schütz, Dr. W. Lesslauer, and Dr. M. Brockhaus for their generous gifts of pBLCAT5, pUChTNFR55, and htr-13 antibody, respectively.

V. Vandevoorde is a Research Assistant with the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen and acknowledges the support from "Kom op tegen kanker." G. Haegeman is a Research Director with the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen. Research was supported by the Interuniversitaire Attractiepolen, the Fonds voor Geneeskundig Wetenschappelijk Onderzoek, and the Vlaams Interuniversitair Instituut voor Biotechnologie, as well as by an EC Biotech Program grant.

Address all correspondence to W. Fiers, Laboratory of Molecular Biology, Flanders Interuniversity Institute for Biotechnology and University of Ghent, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium. Tel.: 32 9 264 51 31. Fax: 32 9 264 53 48.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



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