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A study of cellular localization of HRS and other endosome markers during spermatogenesis in Drosophila melanogaster using chimeric GFP constructs

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Abstract

Acrosome is a specialized organelle in spermatozoids necessary for fertilization of oocyte. Acrosome is formed, according to various theories, either from vesicles of the Golgi apparatus or from endosomes and lysosomes. Hepatocyte growth factor regulated tyrosine kinase substrate (Hrs) is a multidomain protein component of ESCRT-0 complex, that participates in sorting of proteins absorbed during endocytosis. It has been shown that, in mammals, Hgs protein (Hrs homologue) possesses the ability to bind the acrosome, but whether Drosophila Hrs has a similar capacity has not been determined. We found that two forms of Hrs protein are expressed in Drosophila testes: a long form, Hrs-B, and a truncated form, Hrs-A. The later lacks a VHS domain and a portion of FYVE domain, both directly involved in Hrs anchoring to endosomes. We also determined that, unlike mammal Hgs, Drosophila Hrs-B isoform and an almost identical truncated form, Hrs8 ([290–760]), are localized not to acrosome, but to spermatocyte fusomes. This localization requires a portion of the protein molecule located between amino acid residues 383 and 472, which presumably correspond to NF2- and/or Stam-binding domains. In situ hybridization for hrs mRNA showed that the gene is expressed during early spermatogenesis. This is consistent with our data that the Hrs protein binds to spermatocyte fusomes and is absent during later spermatogenesis stages. In addition, we have shown that Hrs is not involved in regulation or implementation of cytokinesis in spermatocytes. Finally, despite the absence of Hrs on Drosophila acrosomes, we detected endosome markers Rab4, Rab7, and Rab11 on this organelle. Thus, our data support the endosomal hypothesis of acrosome biogenesis.

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Abbreviations

MVB:

multivesicular bodies

ESCRT:

endosomal sorting complex required for transport

STAM:

signal-transducing adaptor molecule

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Authors and Affiliations

  1. Institute of Molecular and Cell Biology Science, Siberian Branch, Russian Academy of Sciences, Novosibirsk, 630090, Russia

    E. V. Marilovtseva, T. D. Dubatolova, Y. A. Galimova, S. A. Kopyl & L. V. Omelyanchuk

  2. Novosibirsk State University, Novosibirsk, 630090, Russia

    L. V. Omelyanchuk

Authors
  1. E. V. Marilovtseva
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  2. T. D. Dubatolova
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  3. Y. A. Galimova
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  4. S. A. Kopyl
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  5. L. V. Omelyanchuk
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Correspondence to L. V. Omelyanchuk.

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Original Russian Text © E.V. Marilovtseva, T.D. Dubatolova, Ya. Galimova, S.A. Kopyl, L.V. Omelyanchuk, 2015, published in Tsitologiya, 2015, Vol. 57, No. 7, pp. 509–517.

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Marilovtseva, E.V., Dubatolova, T.D., Galimova, Y.A. et al. A study of cellular localization of HRS and other endosome markers during spermatogenesis in Drosophila melanogaster using chimeric GFP constructs. Cell Tiss. Biol. 9, 452–461 (2015). https://doi.org/10.1134/S1990519X15060073

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  • DOI: https://doi.org/10.1134/S1990519X15060073

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