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Standardized Methods for Measuring Induction of the Heat Shock Response in Caenorhabditis elegans
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Heat shock and developmental expression of hsp83 in the filarial nematode Brugia pahangi

F J Thompson1, A C Cockroft, I Wheatley

  • 1Department of Veterinary Parasitology, University of Glasgow, Scotland, UK.

European Journal of Biochemistry
|November 28, 2001
PubMed

Insights

The Brugia pahangi hsp83 gene is expressed in mammalian-dwelling stages. Its expression requires a temperature threshold of at least 35°C, suggesting a role in thermotolerance for filarial parasites.

Area of Science:

  • Molecular biology
  • Parasitology
  • Heat shock proteins

Background:

  • The heat shock protein 83 (hsp83) is crucial for cellular function under stress.
  • Filarial nematodes like Brugia pahangi inhabit mammalian hosts, experiencing relatively stable temperatures.

Purpose of the Study:

  • To characterize the expression pattern and regulation of the Brugia pahangi hsp83 (Bp-hsp83) gene.
  • To determine the temperature threshold for Bp-hsp83 activation.

Main Methods:

  • Cloning and sequencing of the Bp-hsp83 gene and its 5' upstream region.
  • Transfection of COS-7 cells and Caenorhabditis elegans with a chloramphenicol acetyltransferase (CAT) reporter construct linked to Bp-hsp83.
  • Analysis of CAT activity under varying temperature conditions.

Main Results:

  • Bp-hsp83 mRNA was constitutively expressed in microfilariae and adult worms at 37°C.
  • Reporter gene assays showed Bp-hsp83 expression in COS-7 cells at 37°C, unaffected by heat shock.
  • Significant CAT activity in transfected C. elegans was observed only at temperatures ≥35°C.

Conclusions:

  • Bp-hsp83 exhibits a high temperature threshold for expression, suggesting its role in thermotolerance at host temperatures.
  • The findings provide insights into the molecular mechanisms of parasite adaptation to mammalian environments.

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