Expression of the small heat shock protein gene, hsp30, in Rana catesbeiana fibroblasts.
In the present study, we examined the expression of the Rana catesbeiana small heat shock protein gene, hsp30, in an FT fibroblast cell line. Northern and western blot analyses revealed that hsp30 mRNA or HSP30 protein was not present constitutively but was strongly induced at a heat shock temperature of 35 degrees C. However, treatment of FT cells with sodium arsenite at concentrations that induced hsp gene expression in other amphibian systems caused cell death. Non-lethal concentrations of sodium arsenite (10 microM) induced only minimal accumulation of hsp30 mRNA or protein after 12 h. Immunocytochemical analyses employing laser scanning confocal microscopy detected the presence of heat-inducible HSP30, in a granular or punctate pattern. HSP30 was enriched in the nucleus with more diffuse localization in the cytoplasm. The nuclear localization of HSP30 was more prominent with continuous heat shock. These heat treatments did not alter FT cell shape or disrupt actin cytoskeletal organization. Also, HSP30 did not co-localize with the actin cytoskeleton.
Mulligan-Tuttle
Anne
A
Department of Biology, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
Heikkila
John J
JJ
D000199:Actins / Q000378:metabolism; D000818:Animals; D018053:Arsenites / Q000494:pharmacology; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D050890:HSP30 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D012333:RNA, Messenger / Q000378:metabolism; D011892:Rana catesbeiana / Q000033:anatomy & histology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017670:Sodium Compounds / Q000494:pharmacology; D013997:Time Factors; D014059:Tongue / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection; D015854:Up-Regulation
D000199:Actins; D018053:Arsenites; D050890:HSP30 Heat-Shock Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D017670:Sodium Compounds; C017947:sodium arsenite