The expression of heat shock protein HSP60A reveals a dynamic mitochondrial pattern in Drosophila melanogaster embryos.
The evolutionarily conserved hsp60 ( heat-shock protein 60) family of molecular chaperones ensures the correct folding of nuclear-encoded proteins after their translocation across the mitochondrial membrane during development as well as after heat-shock treatment. Although the overexpression of HSP60 proteins and their localization in the cytoplasm have been linked with many humans pathologies, the detailed pattern of their expression in different animal models and their subcellular localization during normal development and in stress conditions are little-known. In this report, we have used two-dimensional gel electrophoresis followed by MALDI-TOF to identify and purify heat shock protein HSP60A of Drosophila melanoagaster. We demonstrate that it is heat-shock inducible and describe two novel antisera, specifically designed to recognize the denatured and native polypeptide, respectively, in Drosophila. Immunoelectron microscopy and immunostaining of Drosophila cells with these antibodies reveals that HSP60A is always localized to the inner membrane of mitochondria. Expression of HSP60A is post-transcriptionally regulated in a highly dynamic pattern during embryogenesis, even under heat-shock conditions. In contrast, in very stressful situations, its expression is upregulated transcriptionally over the entire embryo. These findings suggest novel roles for HSP60 family proteins during normal Drosophila development.
Baena-López
Luis Alberto
LA
Centro de Biología Molecular "Severo Ochoa", CSIC-UAM, Universidad Autónoma de Madrid, Madrid, Spain.
Alonso
Jana
J
Rodriguez
Javier
J
Santarén
Juan F
JF
D000818:Animals; D018834:Chaperonin 60 / Q000096:biosynthesis* / Q000235:genetics; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006031:Glycosylation; D007150:Immunohistochemistry; D008928:Mitochondria / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018834:Chaperonin 60