Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1997 Jun 10;94(12):6346-51.
doi: 10.1073/pnas.94.12.6346.

CD27, a member of the tumor necrosis factor receptor family, induces apoptosis and binds to Siva, a proapoptotic protein

K V Prasad  1 , Z Ao, Y Yoon, M X Wu, M Rizk, S Jacquot, S F Schlossman
Affiliations

CD27, a member of the tumor necrosis factor receptor family, induces apoptosis and binds to Siva, a proapoptotic protein

K V Prasad et al. Proc Natl Acad Sci U S A. .

Abstract

Members of the tumor necrosis factor receptor (TNFR) superfamily are important for cell growth and survival. In addition to providing costimulatory signals for cell proliferation, ligation of both TNFR1 and Fas can result in programmed cell death or apoptosis. The underlying mechanism requires an intact 80-aa stretch present in the cytoplasmic tails of both TNFR1 and Fas, termed the death domain (DD). Here we show that CD27, a member of the TNFR family, expressed on discrete subpopulations of T and B cells and known to provide costimulatory signals for T and B cell proliferation and B cell Ig production, can also induce apoptosis. Co-crosslinking of surface Ig receptors along with ligation of CD27 augments CD27-mediated apoptosis. Unlike TNFR1 and Fas, the cytoplasmic tail of CD27 is relatively short and lacks the DD. Using the yeast two-hybrid system, we have cloned a novel protein (Siva) that binds to the CD27 cytoplasmic tail. It has a DD homology region, a box-B-like ring finger, and a zinc finger-like domain. Overexpression of Siva in various cell lines induces apoptosis, suggesting an important role for Siva in the CD27-transduced apoptotic pathway.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Ligation of CD27 induces apoptosis. (A) Ramos cells (0.5 × 106/ml) were cultured with increasing concentrations (0.1, 0.2, and 0.5 × 106/ml) of mock (lanes 2–4) and CD70 transfectants (lanes 5–7) for 2 days. Ramos, mock, and CD70 transfectants when cultured alone are shown (lanes 8–10). (B–E) Experiments with formaldehyde-fixed CD70 and mock transfectants, using Ramos, Raji, human CD27 transfected mouse pre-B 300 cell lines and human peripheral blood lymphocytes preactivated with phytohemagglutinin/IL-2. The effect of coligation of surface IgM (Ramos and Raji) and Ig (murine pre-B 300) using anti-human IgM and anti-mouse IgG antibodies (10 μg/ml), respectively, on CD27-induced apoptosis is also shown (B, lanes 7 and 4; C, lane 6 vs. lane 3; and D, lane 5 vs. lane 3).
Figure 2
Figure 2
(A) Siva cDNA sequence obtained from HeLa and human thymus cDNA libraries with the translated ORF (189 aa) is shown. The initiation codon is defined by the upstream in-frame stop codon (−42). The coding region is followed by a stop codon (568), a polyadenylylation signal (709–714). The sequence has been deposited in the GenBank under the accession number U2938. (B) A death domain (DD) homology region toward the amino terminus has been identified in Siva, and its homology to FADD and RIP DDs was determined with use of the multiple sequence alignment program clustal. The shaded regions show homology and the residues in boldface type depict identical residues conserved among the three regions. (C) Putative B-box domain of Siva. Homologous residues between Siva and other proteins in the B-box domain are shaded.
Figure 3
Figure 3
(A) mRNA expression of Siva by Northern blot analysis is shown (upper bands) and is compared with the expression of G3PDH (lower bands). (B) CD27 binds to Siva in 293 cells. CD27WT DNA was transiently coexpressed with either GFP or GFP–Siva for 2 days. The CD27 receptor expression appeared to be similar as seen by flow cytometric analysis (data not shown). CD27 immunoprecipitates using anti-CD27 antibody (IA4) were separated by SDS/PAGE, transferred, and blotted with anti-GFP antiserum. Similar amounts of GFP and GFP–Siva were expressed as seen in the lanes corresponding to whole-cell lysates (lanes 4 and 3). CD27 coprecipitaed GFP–Siva (lane 1) but not GFP (lane 2). (C) Siva-induced apoptosis. Jurkat, Raji, SKW, Ramos, and 293 cells were transiently transfected (2 days) with GFP (hatched bar) and GFP–Siva (solid bar). The histograms represent the percentage of dead cells (based on cell morphology) in the cells expressing green fluorescence. In the case of 293 cells, along with GFP and GFP–Siva, pSVb containing the β-galactosidase gene was cotransfected. β-Galactosidase expression was visualized by X-Gal. The percentage of rounded shrunken cells in the fluorescent population of cells is presented. (D) Transient expression of GFP–Siva but not GFP can induce apoptosis in 293 cells (lane 3 vs. lane 2), the murine pre-B cell line (lane 6 vs. lane 5), and Raji cells (lane 8 vs. lane 7), as judged by the generation of DNA ladder.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Smith C A, Farrah T, Goodwin R G. Cell. 1994;76:959–962. - PubMed
    1. McDonald N Q, Hendrickson W A. Cell. 1993;73:421–424. - PubMed
    1. Camerini D, Walz G, Loenen W A M, Borst J, Seed B. J Immunol. 1991;147:3165–3169. - PubMed
    1. Gravestein L A, Bloom B, Nolten L A, Van Der Vries E. Eur J Immunol. 1993;23:943–950. - PubMed
    1. Goodwin R G, Alderson M R, Smith C A, Armitage R J, Vanden Bos T, Jerzy R, Tough T W, Schoenborn M J, Davis-Smith T, Hennen K, Falk B, Cosman D, Baker E, Sutherland G R, Grabstein K H, Farrah T, Giri J G, Beckmann P M. Cell. 1993;73:447–456. - PubMed

Publication types

MeSH terms

Substances

Associated data