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Biochemical and immunofluorescence analysis of the constitutively expressed HSP27 stress protein in monkey CV-1 cells

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Abstract

The α-crystallin-related stress protein HSP27, which promotes cellular resistance to different types of stress, is constitutively expressed during the growth of several primate tissue culture cells. Here, we report an analysis of the cellular localization of this protein in CV-1 monkey cells. Following cell lysis and fractionation in the absence of detergent about 2 5 % of the cellular content of HSP27 was recovered in the particu late fractions while the remaining of this protein was in the soluble cytoplasmic fraction. This association of HSP27 with particulate fractions was no more observed when cells were lysed in the presence of non-ionic detergent or when cells were pretreated with drugs, such as monensin and colcemid, that disrupt cytoskeletal architecture. Immunofluorescence analysis revealed that HSP27 is concentrated in a polarized perinuclear zone of CV-1 cells from where microtubules radiate. The particular locale of HSP27 was investigated in cells exposed to drugs or treatments, such as monensin, colcemid, cold stess and serum starvation, that disrupt the cellular architecture of microtubules. A correlation was observed between HSP27 cellular locale and microtubules integrity. Our results suggest a possible interaction of a fraction of HSP27 with cytoplasmic organelles or structures, different from the Golgi apparatus, whose distribution depends upon the organization of microtubules.

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Authors and Affiliations

  1. Laboratoire du stress cellulaire, Université Claude Bernard Lyon-I, Centre de Genetique Moleculaire et Cellulaire, CNRS-UMR 106, 43 Boulevard du 11 Novembre, F-69622, Villeurbanne Cedex, France

    X. Preville, P. Mehlen, N. Fabre-Jonca, S. Chaufour, C. Kretz-Remy, M. R. Michel & A. -P. Arrigo

  2. Institute of Medical Microbiology, University of Bern Medical School, CH 3010, Bern, Switzerland

    M. R. Michel

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  1. X. Preville
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  2. P. Mehlen
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  3. N. Fabre-Jonca
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  4. S. Chaufour
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  5. C. Kretz-Remy
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  6. M. R. Michel
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  7. A. -P. Arrigo
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Correspondence to A. -P. Arrigo.

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Preville, X., Mehlen, P., Fabre-Jonca, N. et al. Biochemical and immunofluorescence analysis of the constitutively expressed HSP27 stress protein in monkey CV-1 cells. J Biosci 21, 221–234 (1996). https://doi.org/10.1007/BF02703110

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