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The CED-4-homologous protein FLASH is involved in Fas-mediated activation of caspase-8 during apoptosis

Nature volume 398, pages 777–785 (1999)Cite this article

An Erratum to this article was published on 01 July 1999

Abstract

Fas is a cell-surface receptor molecule that relays apoptotic (cell death) signals into cells. When Fas is activated by binding of its ligand, the proteolytic protein caspase-8 is recruited to a signalling complex known as DISC by binding to a Fas-associated adapter protein. A large new protein, FLASH, has now been identified by cloning of its complementary DNA. This protein contains a motif with oligomerizing activity whose sequence is similar to that of the Caenorhabditis elegans protein CED-4, and another domain (DRD domain) that interacts with a death-effector domain in caspase-8 or in the adapter protein. Stimulated Fas binds FLASH, so FLASH is probably a component of the DISC signalling complex. Transient expression of FLASH activates caspase-8, whereas overexpression of a truncated form of FLASH containing only one of its DRD or CED-4-like domains does not allow activation of caspase-8 and Fas-mediated apoptosis to occur. Overexpression of full-length FLASH blocks the anti-apoptotic effect of the adenovirus protein E1B19K. FLASH is therefore necessary for the activation of caspase-8 in Fas-mediated apoptosis.

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Figure 1: Structure of mouse FLASH and sequence comparison.
Figure 2: Tissue distribution of mouse FLASH mRNA.
Figure 3: Overexpressed FLASH interacts with caspase-8, FADD and Fas in mammalian cells.
Figure 4: Endogenous FLASH was recruited to activated Fas in mouse WR19L12a and human SKW64 and Jurkat cells.
Figure 5: Effects of transfected FLASH and its mutants on activation of caspase-8 and Fas-induced apoptosis.
Figure 6: Self-association of FLASH through its CED-4-homologous domain.
Figure 7: FLASH attenuates the anti-apoptotic effect of adenovirus protein E1B19K.

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Acknowledgements

We thank Y. Tsujimoto for cDNAs encoding Bcl-2 and SKW6.4 cells, D. V. Goeddel for mouse FADD cDNA, D. J. Pickup for CrmA cDNA, S. Hashimoto for antiserum against E1B19K, S.Tsukumo for pME18S-Flag2, H. Kazama for pME18S–Myc and FH2 cells, and Y. Nakanishi for HF1 cells. This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan, and by the Ministry of Health and Welfare, Japan.

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  1. Institute for Virus Research, Kyoto University, Shogoin, Sakyo-ku, 606-8507, Kyoto, Japan

    Yuzuru Imai, Takaharu Kimura, Akira Murakami, Nobuyuki Yajima, Kazuhiro Sakamaki & Shin Yonehara

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Correspondence to Shin Yonehara.

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Imai, Y., Kimura, T., Murakami, A. et al. The CED-4-homologous protein FLASH is involved in Fas-mediated activation of caspase-8 during apoptosis. Nature 398, 777–785 (1999). https://doi.org/10.1038/19709

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