Reovirus-induced apoptosis is preceded by increased cellular calpain activity and is blocked by calpain inhibitors

被引:83
作者
Debiasi, RL
Squier, MKT
Pike, B
Wynes, M
Dermody, TS
Cohen, JJ
Tyler, KL
机构
[1] Univ Colorado, Hlth Sci Ctr, Dept Neurol, Denver, CO 80262 USA
[2] Univ Colorado, Hlth Sci Ctr, Dept Pediat Infect Dis, Denver, CO 80262 USA
[3] Univ Colorado, Hlth Sci Ctr, Dept Med, Denver, CO 80262 USA
[4] Univ Colorado, Hlth Sci Ctr, Dept Microbiol, Denver, CO 80262 USA
[5] Univ Colorado, Hlth Sci Ctr, Dept Immunol, Denver, CO 80262 USA
[6] Denver Vet Affairs Med Ctr, Denver, CO 80262 USA
[7] Vanderbilt Univ, Sch Med, Dept Pediat, Nashville, TN 37232 USA
[8] Vanderbilt Univ, Sch Med, Dept Microbiol & Immunol, Nashville, TN 37232 USA
[9] Vanderbilt Univ, Sch Med, Elizabeth B Lamb Ctr Pediat Res, Nashville, TN 37232 USA
关键词
D O I
10.1128/JVI.73.1.695-701.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The cellular pathways of apoptosis have not been fully characterized; however, calpain, a cytosolic calcium-activated cysteine protease, has been implicated in several forms of programmed cell death. Reoviruses induce apoptosis both in vitro and in vivo and serve as a model for studying virus-induced cell death. We investigated the potential role of calpain in reovirus-induced apoptosis in vitro by measuring calpain activity as well as evaluating the effects of calpain inhibitors, L929 cells were infected with reovirus type 3 Abney (T3A), and calpain activity, measured as cleavage of the fluorogenic calpain substrate Suc-Leu-Leu-Val-Tyr-AMC, was monitored. There was a 1.6-fold increase in calpain activity in T3A-infected cells compared to mock-infected cells; this increase was completely inhibited by preincubation with calpain inhibitor I (N-acetyl-leucyl-leucyl-norleucinal [aLLN]), an active site inhibitor. Both aLLN and PD150606, a specific calpain inhibitor that interacts with the calcium-binding site, inhibited reovirus-induced apoptosis in L929 cells by 54 to 93%. Apoptosis induced by UV-inactivated reovirus was also reduced 65 to 69% by aLLN, indicating that inhibition of apoptosis by calpain inhibitors is independent of effects on viral replication. We conclude that calpain activation is a component of the regulatory cascade in reovirus-induced apoptosis.
引用
收藏
页码:695 / 701
页数:7
相关论文
共 78 条
  • [31] PROTEASE ACTIVATION DURING APOPTOSIS - DEATH BY 1000 CUTS
    MARTIN, SJ
    GREEN, DR
    [J]. CELL, 1995, 82 (03) : 349 - 352
  • [32] APOPTOSIS AND THE CELL-CYCLE
    MEIKRANTZ, W
    SCHLEGEL, R
    [J]. JOURNAL OF CELLULAR BIOCHEMISTRY, 1995, 58 (02) : 160 - 174
  • [33] CALPAIN AND CALPASTATIN
    MURACHI, T
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1983, 8 (05) : 167 - 169
  • [34] Apoptosis in development and disease of the nervous system .1. Naturally occurring cell death in the developing nervous system
    Narayanan, V
    [J]. PEDIATRIC NEUROLOGY, 1997, 16 (01) : 9 - 13
  • [35] Effects of ICE-like protease and calpain inhibitors on neuronal apoptosis
    Nath, R
    Raser, KJ
    McGinnis, K
    Nadimpalli, R
    Stafford, D
    Wang, KKW
    [J]. NEUROREPORT, 1996, 8 (01) : 249 - 255
  • [36] NATH R, 1996, BIOCHEM J, V319, P686
  • [37] Neuberger T, 1997, J NEUROSCI RES, V47, P521, DOI 10.1002/(SICI)1097-4547(19970301)47:5<521::AID-JNR7>3.3.CO
  • [38] 2-M
  • [39] Reovirus infection and tissue injury in the mouse central nervous system are associated with apoptosis
    Oberhaus, SM
    Smith, RL
    Clayton, GH
    Dermody, TS
    Tyler, KL
    [J]. JOURNAL OF VIROLOGY, 1997, 71 (03) : 2100 - 2106
  • [40] Oberhaus SM, 1998, CURR TOP MICROBIOL, V233, P23