Expression of heat shock protein 70 in the thermally stressed Antarctic clam Laternula elliptica

Abstract

Heat shock protein 70 (designated Laternula elliptica Hsp70 (LEHsp70)) expression was investigated in an Antarctic mud clam to see whether or not the inducible heat shock response has been conserved throughout over 25 million years of adaptation to constant low environmental temperatures. LEHsp70 cDNA was cloned and sequenced from the Antarctic clam Latemula elliptica. We used degenerated primers designed in the highly conserved regions of Hsp to amplify the corresponding mRNA, and full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full length of LEHsp70 cDNA was 2470 bp, with a 5' untranslated region (UTR) of 92 bp, a 3' UTR of 416 bp, and an open reading frame (ORF) of 1962 bp encoding a polypeptide of 653 amino acids with an estimated molecular mass of 71.266 kDa and an estimated isoelectric point of 5.20. LEHsp70 contained highly conserved functional motifs of the cytosolic Hsp70 family. Expression of the LEHsp70 gene was quantified by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) of digestive gland and gill tissues. Heat shock (10 degrees C for different time periods) caused rapid induction of LEHsp70. A significant 4.6 +/- 0.14-fold increase in the LEHsp70/beta-Actin mRNA ratio occurred in the gill at 12 hours, which returned to baseline after 48 hours. In contrast, the maximum expression in the digestive gland (3.6 +/- 0.36) was reached at 24 hours and was still significant after 48 hours (1.89 +/- 0.21). This indicates that LEHsp70 may play an important role in mediating thermal stress and tolerance in this clam.