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Permeability changes resulting from virus-cell fusion: temperature-dependence of the contributing processes

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Summary

A new assay for membrane fusion, using the fluorescent probe pyrene-sulphonyl-phosphatidyl ethanolamine, has been developed. Fusion between the envelope of Sendai virus and human erythrocytes or Lettre cells has a Q10 of ∼4 at 37° C, increasing to ∼7 at 7 ° C; there is no lag to onset of fusion. Viral neuraminidase has a Q10 of 2.3 between 37° C and 4° C. Its action limits the extent of fusion by causing the elution of virus; this effect is particularly marked at low temperature because of the difference in Q10 for fusion and neuraminidase. The temperature-dependence of the initiation of permeability changes following the removal of inhibitory amounts of Ca2+ is ∼2; thus membrane fusion is the principal temperature-sensitive step during the permeabilization of cells by Sendai virus. A recovery process, by which cells become insensitive to the removal of Ca2+ and which therefore limits the extent of permeabilization, has a Q10 of 7.4 between 37° C and 21° C. It is concluded that the lag to onset of permeability changes is not due to a lag in virus-cell membrane fusion, but to the gradual acquisition of a threshold level of membrane damage; the extent of permeabilization depends on the rate of fusion relative to the rates of neuraminidase and recovery.

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  1. K. J. Micklem

    Present address: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, UK

Authors and Affiliations

  1. Department of Biochemistry, St George's Hospital Medical School, SW17 ORE, Cranmer Terrace, London, UK

    K. J. Micklem, A. Nyaruwe & C. A. Pasternak

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  3. C. A. Pasternak
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Micklem, K.J., Nyaruwe, A. & Pasternak, C.A. Permeability changes resulting from virus-cell fusion: temperature-dependence of the contributing processes. Mol Cell Biochem 66, 163–173 (1985). https://doi.org/10.1007/BF00220784

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