Apoptosis of T cells mediated by Ca2+-induced release of the transcription factor MEF2

Science

Volumn 286, Issue 5440, 1999, Pages 790-793

  Youn, Hong Duk ^a   Sun, Luo ^a   Prywes, Ron ^b   Liu, Jun O ^a  

^a MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

^b Och Spine at New York Presbyterian Hospitals   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CALCINEURIN; CALCIUM; CALMODULIN; STEROID RECEPTOR; T LYMPHOCYTE RECEPTOR; TRANSCRIPTION FACTOR;

APOPTOSIS; ARTICLE; CALCIUM CELL LEVEL; HUMAN; HUMAN CELL; PRIORITY JOURNAL; PROTEIN BINDING; PROTEIN EXPRESSION; SIGNAL TRANSDUCTION; T LYMPHOCYTE; THYMOCYTE;

APOPTOSIS; CALCINEURIN; CALCIUM; CALCIUM SIGNALING; CALMODULIN; CELL LINE; DNA-BINDING PROTEINS; GENE EXPRESSION; GENES, REPORTER; HUMANS; JURKAT CELLS; MYOGENIC REGULATORY FACTORS; PHOSPHOPROTEINS; RECEPTORS, ANTIGEN, T-CELL; RECEPTORS, CYTOPLASMIC AND NUCLEAR; RECEPTORS, STEROID; T-LYMPHOCYTES; TRANSCRIPTION FACTORS; TRANSCRIPTION, GENETIC; TWO-HYBRID SYSTEM TECHNIQUES;

EID: 0033595816     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5440.790     Document Type: Article

References (28)

1. A. Winoto, Curr. Opin. Immunol. 9, 365 (1997).
2. B. Wong and Y. Choi, ibid., p. 358.
3. J. D. Woronicz, B. Calnan, V. Ngo, A. Winoto, Nature 367, 277 (1994).
4. Z. G. Liu, S. W. Smith, K. A. McLaughlin, L. M. Schwartz, B. A. Osborne, ibid., p. 281.
5. L. E. Cheng, F. K. Chan, D. Cado, A. Winoto, EMBO J. 16, 1865 (1997).
6. J. D. Woronicz et al., Mol. Cell. Biol. 115, 6364 (1995).
7. L. A. Gossett, D. J. Kelvin, E. A. Sternberg, E. N. Olson, ibid. 9, 5022 (1989).
8. R. Pollock and R. Treisman, Genes Dev. 5, 2327 (1991).
9. Y. T. Yu et al., ibid. 6, 1783 (1992).
10. D. Leifer et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1546 (1993).
11. J. F. Martin, J. J. Schwarz, E. N. Olson, ibid., p. 5282.
12. J. F. Martin et al., Mol. Cell. Biol. 14, 1647 (1994).
13. L. Sun et al., Immunity 8, 703 (1998).
14. M. M. Lai, P. E. Burnett, H. Wolosker, S. Blackshaw, S. H. Snyder, J. Biol. Chem. 273, 18325 (1998).
15. H.-D. Youn and J. O. Liu, unpublished data.
16. H.-L. Hsu, I. Wadman, R. Baer, Proc. Natl. Acad. Sci. U.S.A. 91, 3181 (1994).
17. H.-D. Youn and J. O. Liu, unpublished data.
18. Web figure 1 can be found at www.sciencemag.org/ feature/data/1039959.shl.
19. M. Ikura et al., Science 256, 632 (1992).
20. B. Corneliussen et al., Nature 368, 760 (1994).
21. 2 or 10 mM EGTA for 1 hour, followed by incubation with CaM-sepharose for 1 hour. The beads were washed four times with Buffer B. Proteins bound to beads were resolved by SDS-polyacrylamide gel electrophoresis (PAGE), transferred to nitrocellulose membrane, and probed using anti-GST-Cabin1 polyclonal antibodies, which recognize both GST and Cabin 1 polypeptides.
22. 2 or 10 mM EGTA. 0.5 μg of GST-Cabin1(2037-2220) was added alone or in combination with 5 μg of CaM to MEF2D-translated products. Reaction mixtures were precipitated using glutathione beads after a 2-hour incubation, separated by 10% SDS-PAGE, and developed by autoradiography.
23. Web figure 2 can be found at www.sciencemag.org/ feature/data/1039959.shl.
24. The pBabeSVCabin1 expression vector was constructed by subcloning the Sal I fragment from pSG-Cabin1, which contains an SV40 promoter and a polyadenylation site flanking the full-length Cabin1 cDNA, into the Sal I site of the pBabe vector [W. S. Pear, G. P. Nolan, M. L. Scott, D. Baltimore, Proc. Natl. Acad. Sci. U.S.A. 90, 8392 (1993)]. DO11.10 cells were transfected with pBabeSVCabin1 and incubated for 24 hours. Transfected cells were treated with puromycin (2 μg/ml) for another 24 hours and dead cells were removed by Ficoll gradient centrifugation. Puromycin-resistant cells were selected by diluting (1:10) transfected cells into puromycin-containing RPMI medium every 3 days until all control DO11.10 cells were dead. The expression of exogenous Cabin1 was confirmed by protein immunoblot using a c-Myc mAb. DNA fragmentation assay was performed as described [K. S. Sellins and J. J. Cohen, J. Immunol. 139, 3199 (1987)].
25. The pBabeSVCabin1 expression vector was constructed by subcloning the Sal I fragment from pSG-Cabin1, which contains an SV40 promoter and a polyadenylation site flanking the full-length Cabin1 cDNA, into the Sal I site of the pBabe vector [W. S. Pear, G. P. Nolan, M. L. Scott, D. Baltimore, Proc. Natl. Acad. Sci. U.S.A. 90, 8392 (1993)]. DO11.10 cells were transfected with pBabeSVCabin1 and incubated for 24 hours. Transfected cells were treated with puromycin (2 μg/ml) for another 24 hours and dead cells were removed by Ficoll gradient centrifugation. Puromycin-resistant cells were selected by diluting (1:10) transfected cells into puromycin-containing RPMI medium every 3 days until all control DO11.10 cells were dead. The expression of exogenous Cabin1 was confirmed by protein immunoblot using a c-Myc mAb. DNA fragmentation assay was performed as described [K. S. Sellins and J. J. Cohen, J. Immunol. 139, 3199 (1987)].
26. B. L. Black and E. N. Olsen, Annu. Rev. Cell Dev. Biol. 14, 167 (1998).
27. H.-D. Youn, T. Chatila, J. O. Liu, unpublished data.
28. We thank A. Winoto, S. Burakoff, A. Lassar, L. Mathews, E. Seto, and L. Lau for reagents; B. Turk, E. Griffith, A. Mondragon, and other members of the Liu lab for helpful suggestions and critical comments; D. Baltimore and associates for access to a luminometer; and R. Hynes for access to a Gene Pulser. Supported in part by National Institute of General Medical Sciences grant GM55783, the Rita Allen Foundation, and Chiba Prefecture, Japan (J.O.L.) and by a NIH training grant (L.S.).