Molecular cloning and characterization of Hsp27.6: the first reported small heat shock protein from Apis cerana cerana

Abstract

Small heat shock proteins (sHSPs) play an important role in the cellular defense of prokaryotic and eukaryotic organisms against a variety of internal and external stressors. In this study, a cDNA clone encoding a member of the α-crystallin/sHSP family, termed AccHsp27.6, was isolated from Apis cerana cerana. The full-length cDNA is 1,014 bp in length and contains a 708-bp open reading frame encoding a protein of 236 amino acids with a calculated molecular weight of 27.6 kDa and an isoelectric point of 7.53. Seven putative heat shock elements and three NF-κB binding sites were present in the 5′-flanking region, suggesting a possible function in immunity. A semi-quantitative RT-PCR analysis indicated that AccHsp27.6 was expressed in all tested tissues and at different developmental stages. Furthermore, expression of the AccHsp27.6 transcript was induced by exposure to heat shock, H 2 O 2 , a number of different chemicals (including SO 2 , formaldehyde, alcohol, acetone, chloroform, and the pesticides phoxime and acetamiprid), and the microbes Staphylococcus aureus and Micrococcus luteus. In contrast, the mRNA expression could be repressed by CO 2 , the pesticides pyriproxyfen and cyhalothrin, and the microbes Bacillus subtilis and Pseudomonas aeruginosa. Notably, the recombinant AccHsp27.6 protein exhibited significant in vitro molecular chaperone activity and antimicrobial activity. Taken together, these results suggest that AccHsp27.6 might play an important role in the response to abiotic and biotic stresses and in immune reactions.

Characterization of the small heat shock protein Hsp27 gene in Chironomus riparius (Diptera) and its expression profile in response to temperature changes and xenobiotic exposures

“…Yet, nonylphenol was able to induce Hsp70 gene activity in C. riparius (Lee and Choi 2006;Morales et al 2011). The differential sensitivity of Hsp27 gene to specific chemicals was also reported in Apis cerana; while only some pesticides were able to upregulate this gene others did not alter it or even downregulate transcription (Liu et al 2012). In contrast, exposure to triclosan, bisphenol A and, particularly, cadmium significantly increased Hsp27 mRNA levels in C. riparius.…”

Characterization of six small HSP genes from Chironomus riparius (Diptera, Chironomidae): Differential expression under conditions of normal growth and heat-induced stress

“…Several sHSPs are known in insects, with a variable number of members appearing in different species. Drosophila melanogaster was the first insect in which they were described (Southgate et al, 1983), though the sHSPs have also been identified and described in lepidopterans such as Bombyx mori (Sakano et al, 2006;Li et al, 2009;Wang et al, 2014), and hymenopterans such Apis cerana cerana (Liu et al, 2012Zhang et al, 2014). However, in most insect species exists only scattered information and a few members identified.…”

Small heat shock proteins, phylogeny in filamentous fungi and expression analyses in Aspergillus nidulans

“…Noteworthily, heat stimulus caused expression of AN7892, AN5781, AN3555 and AN2530 increased by more than 40-fold at all of the time points tested, while mRNA levels of these four sHSPs were only up-regulated by 6.3-to 41.4-fold in the cold condition(4°C), indicating that thermal shock induction was more intensive than cold shock, which was consistent with the recent studies of sHSPs in P. xylostella (Chen and Zhang, 2014). Osmotic and oxidative stresses may also trigger expression of shsp genes, e.g., all nine shsp genes in T. hispida were upregulated at least one stress time-point under NaCl stress, while overproduction of sHSPs, for example, HSP26.6 in Oryza sativa, HSP17.7 in Arabidopsis thaliana and HSP 27.6 in Apis cerana, was observed in response to H 2 O 2 treatment (Sun et al, 2002;Liu et al, 2012). In Ustilago maydis, an increased expression of three shsp genes (hsp12, hsp20 and hsp30) was also revealed in response to oxidative stress (Ghosh, 2014).…”