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Volumn 278, Issue 5336, 1997, Pages 294-298

Caspase-3-generated fragment of gelsolin: Effector of morphological change in apoptosis

Author keywords

[No Author keywords available]

Indexed keywords

CALCIUM; COMPLEMENTARY DNA; CYSTEINE PROTEINASE; DNA FRAGMENT; FAS ANTIGEN; GELSOLIN;

EID: 0030820875     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5336.294     Document Type: Article
Times cited : (1061)

References (44)
  • 1
    • 0027973061 scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1994) J. Biol. Chem. , vol.269 , pp. 30761
    • Fernandes-Alnemri, T.1    Litwack, G.2    Alnemri, E.S.3
  • 2
    • 0028990125 scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1995) Cell , vol.81 , pp. 801
    • Tewari, M.1
  • 3
    • 0029956641 scopus 로고    scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1996) Nature , vol.384 , pp. 368
    • Kuida, K.1
  • 4
    • 0009530192 scopus 로고    scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1996) Cell , vol.87 , pp. 171
    • Alnemri, E.S.1
  • 5
    • 0025719085 scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1991) Annu. Rev. Cell Biol. , vol.7 , pp. 663
    • Ellis, R.E.1    Yuan, J.Y.2    Horvitz, H.R.3
  • 6
    • 0029880987 scopus 로고    scopus 로고
    • T. Fernandes-Alnemri, G. Litwack, E. S. Alnemri, J. Biol. Chem. 269, 30761 (1994); M. Tewari et al., Cell 81, 801 (1995); K. Kuida et al., Nature 384, 368 (1996); E. S. Alnemri et al., Cell 87, 171 (1996); R. E. Ellis, J. Y. Yuan, H. R. Horvitz, Annu. Rev. Cell Biol. 7, 663 (1991); D. Xue, S. Shaham, H. R. Horvitz, Genes Dev. 10, 1073 (1996).
    • (1996) Genes Dev. , vol.10 , pp. 1073
    • Xue, D.1    Shaham, S.2    Horvitz, H.R.3
  • 10
    • 0031149841 scopus 로고    scopus 로고
    • R. W. King, K. D. Lustig, P. T. Stukenberg, T. J. McGarry, M. W. Kirschner, Science 277, 973 (1997); P. T. Stukenberg et al., Curr. Biol. 7, 338 (1997).
    • (1997) Curr. Biol. , vol.7 , pp. 338
    • Stukenberg, P.T.1
  • 12
    • 1842318496 scopus 로고    scopus 로고
    • note
    • 35S-labeled gelsolin. Cleavage of gelsolin was monitored by SDS-PAGE and autoradiography and occurred concurrently with caspase-3 - like activity on DEVD - 7-amino-4-methylcoumarin.
  • 13
    • 0022487183 scopus 로고
    • D. J. Kwiatkowski et al., Nature 323, 455 (1986); M. Way and A. Weeds, J. Mol. Biol. 203, 1127 (1988); C. W. Dieffenbach, D. N. Sen Gupta, D. Krause, D. Sawzak, R. H. Silverman, J. Biol. Chem. 264, 13281 (1989).
    • (1986) Nature , vol.323 , pp. 455
    • Kwiatkowski, D.J.1
  • 14
    • 0024293322 scopus 로고
    • D. J. Kwiatkowski et al., Nature 323, 455 (1986); M. Way and A. Weeds, J. Mol. Biol. 203, 1127 (1988); C. W. Dieffenbach, D. N. Sen Gupta, D. Krause, D. Sawzak, R. H. Silverman, J. Biol. Chem. 264, 13281 (1989).
    • (1988) J. Mol. Biol. , vol.203 , pp. 1127
    • Way, M.1    Weeds, A.2
  • 16
    • 1842349240 scopus 로고    scopus 로고
    • note
    • Native and cleaved gelsolin preparations were analyzed on a BioSil SEC250 column, run in 180 mM NaCI with 10 mM phosphate buffer (pH 7.4) at a flow rate of 1 ml/min, and monitored at 280 nm. Column fractions were analyzed by SDS-PAGE and Coomassie staining.
  • 20
    • 0027326858 scopus 로고
    • W. C. Liles, P. A. Kiener, J. A. Ledbetter, A. Aruffo, S. J. Klebanoff, J. Exp. Med. 184, 429 (1996); Y. Takeda et al., Int. Immunol. 5, 691 (1993).
    • (1993) Int. Immunol. , vol.5 , pp. 691
    • Takeda, Y.1
  • 23
    • 1842308972 scopus 로고    scopus 로고
    • note
    • 6) were fixed with 4% paraformaldehyde and permeabilized with ethanol/acetone. After washing with phosphate-buffered saline (PBS), cells were incubated with TdT and fluorescein-conjugated dUTP at 37°C for 1 hour. Cells were washed, and free DNA ends were visualized by fluorescent microscopy.
  • 25
    • 0028914814 scopus 로고
    • W. Witke et al., Cell 81, 41 (1995).
    • (1995) Cell , vol.81 , pp. 41
    • Witke, W.1
  • 26
    • 1842396785 scopus 로고    scopus 로고
    • note
    • DNA was prepared from cells by standard digestion and organic extraction. Extracted DNA (10 μg) was separated by electrophoresis in 1.5% agarose and visualized with ethidium staining.
  • 27
    • 1842392867 scopus 로고    scopus 로고
    • note
    • 5) were treated with TNF+CHX or antibody to Fas, rinsed with PBS, and incubated in 50 mM tris-HCl (pH 7.4), 1 mM EDTA, 10 mM EGTA, and 10 μM digitonin at 37°C for 10 min. After centrifugation at 15,000g for 10 min, the lysate was incubated in DEVD-AFC (Enzyme Systems Products) for 30 min. Released AFC was measured by spectrofluorimetry with excitation at a wavelength of 380 nm and emission at 460 nm.
  • 29
    • 0029610432 scopus 로고
    • T. Mashima, M. Naito, N. Fujita, K. Noguchi, T. Tsuruo, Biochem. Biophys. Res. Commun. 217, 1185 (1995); C. Kayalar, T. Ord, M. P. Testa, L. T. Zhong, D. E. Bredesen, Proc. Natl. Acad. Sci. U.S.A. 93, 2234 (1996); Q. Song et al., ibid. 94, 157 (1997); C. Brancolini, M. Benedetti, C. Schneider, EMBO J. 14, 5179 (1995).
    • (1995) Biochem. Biophys. Res. Commun. , vol.217 , pp. 1185
    • Mashima, T.1    Naito, M.2    Fujita, N.3    Noguchi, K.4    Tsuruo, T.5
  • 30
    • 0029876247 scopus 로고    scopus 로고
    • T. Mashima, M. Naito, N. Fujita, K. Noguchi, T. Tsuruo, Biochem. Biophys. Res. Commun. 217, 1185 (1995); C. Kayalar, T. Ord, M. P. Testa, L. T. Zhong, D. E. Bredesen, Proc. Natl. Acad. Sci. U.S.A. 93, 2234 (1996); Q. Song et al., ibid. 94, 157 (1997); C. Brancolini, M. Benedetti, C. Schneider, EMBO J. 14, 5179 (1995).
    • (1996) Proc. Natl. Acad. Sci. U.S.A. , vol.93 , pp. 2234
    • Kayalar, C.1    Ord, T.2    Testa, M.P.3    Zhong, L.T.4    Bredesen, D.E.5
  • 31
    • 0031024495 scopus 로고    scopus 로고
    • T. Mashima, M. Naito, N. Fujita, K. Noguchi, T. Tsuruo, Biochem. Biophys. Res. Commun. 217, 1185 (1995); C. Kayalar, T. Ord, M. P. Testa, L. T. Zhong, D. E. Bredesen, Proc. Natl. Acad. Sci. U.S.A. 93, 2234 (1996); Q. Song et al., ibid. 94, 157 (1997); C. Brancolini, M. Benedetti, C. Schneider, EMBO J. 14, 5179 (1995).
    • (1997) Proc. Natl. Acad. Sci. U.S.A. , vol.94 , pp. 157
    • Song, Q.1
  • 32
    • 0028788309 scopus 로고
    • T. Mashima, M. Naito, N. Fujita, K. Noguchi, T. Tsuruo, Biochem. Biophys. Res. Commun. 217, 1185 (1995); C. Kayalar, T. Ord, M. P. Testa, L. T. Zhong, D. E. Bredesen, Proc. Natl. Acad. Sci. U.S.A. 93, 2234 (1996); Q. Song et al., ibid. 94, 157 (1997); C. Brancolini, M. Benedetti, C. Schneider, EMBO J. 14, 5179 (1995).
    • (1995) EMBO J. , vol.14 , pp. 5179
    • Brancolini, C.1    Benedetti, M.2    Schneider, C.3
  • 33
    • 0029612323 scopus 로고    scopus 로고
    • D. A. Schafer and J. A. Cooper, Annu. Rev. Cell Dev. Biol. 11, 497 (1995); H. D. Campbell et al., Proc. Natl. Acad. Sci. U.S.A. 90, 11386 (1993); D. J. Kwiatkowski et al., unpublished data.
    • (1995) Annu. Rev. Cell Dev. Biol. , vol.11 , pp. 497
    • Schafer, D.A.1    Cooper, J.A.2
  • 34
    • 0027496510 scopus 로고
    • D. A. Schafer and J. A. Cooper, Annu. Rev. Cell Dev. Biol. 11, 497 (1995); H. D. Campbell et al., Proc. Natl. Acad. Sci. U.S.A. 90, 11386 (1993); D. J. Kwiatkowski et al., unpublished data.
    • (1993) Proc. Natl. Acad. Sci. U.S.A. , vol.90 , pp. 11386
    • Campbell, H.D.1
  • 35
    • 0029612323 scopus 로고    scopus 로고
    • unpublished data
    • D. A. Schafer and J. A. Cooper, Annu. Rev. Cell Dev. Biol. 11, 497 (1995); H. D. Campbell et al., Proc. Natl. Acad. Sci. U.S.A. 90, 11386 (1993); D. J. Kwiatkowski et al., unpublished data.
    • Kwiatkowski, D.J.1
  • 36
    • 0029144456 scopus 로고
    • M. Tanaka et al., Cancer Res. 55, 3228 (1995); H. L. Asch et al., ibid. 56, 4841 (1996); L. Zhang et al., Science 276, 1268 (1997).
    • (1995) Cancer Res. , vol.55 , pp. 3228
    • Tanaka, M.1
  • 37
    • 0029854630 scopus 로고    scopus 로고
    • M. Tanaka et al., Cancer Res. 55, 3228 (1995); H. L. Asch et al., ibid. 56, 4841 (1996); L. Zhang et al., Science 276, 1268 (1997).
    • (1996) Cancer Res. , vol.56 , pp. 4841
    • Asch, H.L.1
  • 38
    • 0030962307 scopus 로고    scopus 로고
    • M. Tanaka et al., Cancer Res. 55, 3228 (1995); H. L. Asch et al., ibid. 56, 4841 (1996); L. Zhang et al., Science 276, 1268 (1997).
    • (1997) Science , vol.276 , pp. 1268
    • Zhang, L.1
  • 39
    • 1842386031 scopus 로고    scopus 로고
    • note
    • 35S]methionine-labeled proteins, using a coupled transcription-translation system (TNT system, Promega). We incubated 2 ml of the translation mix with 0.5 ml of active or inactivated caspase-3 (0.15 ng/ml) at 30°C for 0.5 hour. Caspase-3 was inactivated by incubation with 50 mM DEVD-fmk for 30 min before addition to the translation products. Recombinant active caspase-3 was prepared by expressing the sequence of human caspase-3 (YAMA/CPP32, a gift of V. Dixit) in E. coli (2). Purified caspase-3 (generously provided by A. L. Fear) was fully active when assayed on DEVD - 7-amino-4-methylcoumarin as previously described (3). Caspase-3 was >80% pure, consisting of 17-and 11-kD subunits, as assayed by SDS-PAGE.
  • 42
    • 0030052368 scopus 로고    scopus 로고
    • Rat embryo fibroblasts (REF 52) [T. Joneson, M. A. White, M. H. Wigler, D. Bar-Sagi, Science 271, 810 (1996)] were seeded on acid-treated glass cover slips, and 70% density cells were microinjected with plasmid DNA pGG-NT (encoding residues 1 through 352 of gelsolin and tagged with EYMPME epitope at the COOH-terminus) or pGG-CT (encoding residues 353 through 731 of gelsolin and tagged with EYMPME epitope at the COOH-terminus) at 0.1 μg/ ml in 100 mM Hepes (pH 7.4). A plasmid encoding green fluorescent protein (GFP) (pEGFPN1, Pro-mega) at 1 ng/ml was coinjected as a marker. Cells were fixed in 4% paraformaldehyde/PBS, permeabilized with 0.1% Triton X-100, and stained for the expression of tagged proteins, using a mouse monoclonal antibody to EYMPME (50 μg/ml) and a secondary goat anti-mouse immunoglobulin G coupled to Texas Red (Amersham). Actin filaments were visualized with tetramethyl rhodamine isothiocyanate (TRITC) - Phalloidin (Sigma).
    • (1996) Science , vol.271 , pp. 810
    • Joneson, T.1    White, M.A.2    Wigler, M.H.3    Bar-Sagi, D.4
  • 44
    • 1842357168 scopus 로고    scopus 로고
    • note
    • -/- cells. Supported by Chiron Corporation and NIH grants RO1 HL54188 and P01 HL48743 to D.J.K.


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