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C. elegans mitochondrial factor WAH-1 promotes phosphatidylserine externalization in apoptotic cells through phospholipid scramblase SCRM-1

Nature Cell Biology volume 9, pages 541–549 (2007)Cite this article

Abstract

Externalization of phosphatidylserine, which is normally restricted to the inner leaflet of plasma membrane, is a hallmark of mammalian apoptosis1,2,3,4. It is not known what activates and mediates the phosphatidylserine externalization process in apoptotic cells. Here, we report the development of an annexin V-based phosphatidylserine labelling method and show that a majority of apoptotic germ cells in Caenorhabditis elegans have surface-exposed phosphatidylserine, indicating that phosphatidylserine externalization is a conserved apoptotic event in worms. Importantly, inactivation of the gene encoding either the C. elegans apoptosis-inducing factor (AIF) homologue (WAH-1)5, a mitochondrial apoptogenic factor, or the C. elegans phospholipid scramblase 1 (SCRM-1), a plasma membrane protein, reduces phosphatidylserine exposure on the surface of apoptotic germ cells and compromises cell-corpse engulfment. WAH-1 associates with SCRM-1 and activates its phospholipid scrambling activity in vitro. Thus WAH-1, after its release from mitochondria during apoptosis, promotes plasma membrane phosphatidylserine externalization through its downstream effector, SCRM-1.

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Figure 1: wah-1 promotes phosphatidylserine exposure on the surface of apoptotic germ cells.
Figure 2: wah-1 RNAi affects the clearance of apoptotic cells.
Figure 3: SCRM-1 is a plasma membrane protein important for cell-corpse engulfment.
Figure 4: scrm-1 and wah-1 function in the same pathway to promote phosphatidylserine exposure in apoptotic cells.
Figure 5: WAH-1 interacts specifically with SCRM-1 to activate its phospholipid-scrambling activity.

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Acknowledgements

We thank T. Blumenthal for anti-CstF-64 antibody and Geraldine Seydoux for the pTE5 construct. This research was supported by a Burroughs Wellcome Fund Career Award and a Human Frontier Science Program (HFSP) grant (RGP0016/2005-C) to D.X., a grant from the Ministry of Education, Culture, Sports, Science and Technology of Japan to S.M., and grants from the National Institutes of Health (NIH) to D.X., X.S.X. and Y.S.

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  1. Xiaochen Wang

    Present address: Current address: National Institute of Biological Sciences, Beijing, 102206, China.,

Authors and Affiliations

  1. Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, 80309, CO, USA

    Xiaochen Wang, Chun-Ling Sun, Chonglin Yang, Yong Shi & Ding Xue

  2. McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, 75390, TX, USA

    Jin Wang & Xiao-Song Xie

  3. Department of Physiology, Tokyo Women's Medical University, School of Medicine, and CREST, JST, Tokyo, 162-8666, Japan

    Keiko Gengyo-Ando, Tetsuo Kobayashi & Shohei Mitani

  4. Department of Molecular Biology, Princeton University, Princeton, 08544, NJ, USA

    Lichuan Gu & Yigong Shi

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Contributions

X.C.W. performed most of the experiments. X.C.W. and D.X. designed and interpreted most of the experiments. J.W. and X.S.W. performed the in vitro proteoliposome assays and related data analysis. K.G.A., T.K. and S.M. isolated scrm deletion alleles. L.G., Y.G.S., C.L.S., Y.S. and C.L.Y. contributed to the experiments. X.C.W., X.S.X. and D.X. wrote the paper and others commented on the manuscript.

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Correspondence to Ding Xue.

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The authors declare no competing financial interests.

Supplementary information

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Supplementary Figures S1, S2, S3, S4, S5, Supplementary Methods, Supplementary Tables S1, S2, S3, S4 and References (PDF 5201 kb)

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Wang, X., Wang, J., Gengyo-Ando, K. et al. C. elegans mitochondrial factor WAH-1 promotes phosphatidylserine externalization in apoptotic cells through phospholipid scramblase SCRM-1. Nat Cell Biol 9, 541–549 (2007). https://doi.org/10.1038/ncb1574

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