RESUMO
Small heat shock proteins (sHSPs) are a group of molecular chaperones and play a crucial role in cell response to various stresses. In this study, we cloned and sequenced a small heat shock protein 27 (LcHSP27) cDNA from large yellow croaker, Larimichthys crocea. The full-length cDNA of LcHSP27 was 1227 bp, including a 5'-terminal untranslated region (UTR) of 54 bp, a 3'-terminal UTR of 561 bp and an open reading frame (ORF) of 612 bp encoding a polypeptide of 203 amino acids. Three conserved phosphorylation sites of serine were identified in the deduced LcHSP27 amino acid sequence at positions 15, 91 and 95 respectively, and a typical α-crystallin domain was at positions 96-193. Phylogenetic analysis revealed that LcHSP27 was categorized together with the HSP27 obtained from other fish. And a closer phylogenetic relationship of HSP27 was found with HSP22, then HSP30. Quantitative real-time reverse transcription PCR (qRT-PCR) analysis indicated the strongest expression of LcHSP27 in heart. However, expression of LcHSP27 in other examined tissues including muscle, brain, liver, spleen, kidney, gill, and blood was very weak. The impact of temperature and cadmium (Cd(2+)) stress on LcHSP27 expression was tested in liver and brain. The results showed that the levels of LcHSP27 expression increased significantly after low temperature (19°C) and high temperature (27°C and 31°C) stress both in liver and brain. And low temperature stress induced a higher LcHSP27 expression in liver. More important, LcHSP27 expression showed a dramatic up-regulation after a combined stress of temperature and cadmium (p<0.05). These results reveal that HSP27 may play an important role in the large yellow croaker response to temperature and cadmium stress.
