Characterization, expression profiling, and thermal tolerance analysis of heat shock protein 70 in pine sawyer beetle, Monochamus alternatus hope (Coleoptera: Cerambycidae)

Abstract

Monochamus alternatus Hope (Coleoptera: Cerambycidae) warrants attention as a dominant transmission vector of the pinewood nematode, and it exhibits tolerance to high temperature. Heat shock protein 70 (HSP70) family members, including inducible HSP70 and heat shock cognate protein 70 (HSC70), are major contributors to the molecular chaperone networks of insects under heat stress. In this regard, we specifically cloned and characterized three MaltHSP70s and three MaltHSC70s. Bioinformatics analysis on the deduced amino acid sequences showed these genes, having close genetic relationships with HSP70s of Coleopteran species, collectively shared conserved signature structures and ATPase domains. Subcellular localization prediction revealed the HSP70s of M. alternatus were located not only in the cytoplasm and endoplasmic reticulum but also in the nucleus and mitochondria. The transcript levels of MaltHSP70s and MaltHSC70s in each state were significantly upregulated by exposure to 35–50°C for early 3 h, while MaltHSP70s reached a peak after exposure to 45°C for 2–3 h in contrast to less-upregulated MaltHSC70s. In terms of MaltHSP70s, the expression threshold in females was lower than that in males. Also, both fat bodies and Malpighian tubules were the tissues most sensitive to heat stress in M. alternatus larvae. Lastly, the ATPase activity of recombinant MaltHSP70-2 in vitro remained stable at 25–40°C, and this recombinant availably enhanced the thermotolerance of Escherichia coli. Overall, our findings unraveled HSP70s might be the intrinsic mediators of the strong heat tolerance of M. alternatus due to their stabilized structure and bioactivity.

Functional analysis of small heat shock proteins providing evidence of temperature tolerance in Hyphantria cunea

“…Heat shock proteins were first discovered in Drosophila melanogaster when exposed to high temperature in 1962 and were subsequently divided into four families based on their molecular weight and function: HSP90, HSP70, HSP60 and small heat shock proteins (sHSPs) (Feder & Hofmann, 1999; King & MacRae, 2015; Ritossa, 1962). There have been a number of lines of evidence supporting the view that HSPs can serve as molecular chaperones to protect other cellular proteins and preserve key enzyme functions under environmental stresses such as heat, cold, starvation, oxidative stress and toxicity in insects or other organisms (Burton et al, 1988; Choi et al, 2008; Li et al, 2020; Li et al, 2021; Pan et al, 2018; Xie et al., 2020; Zhang et al., 2014;). The various HSP families form complex chaperone networks that participate in protein synthesis, folding, refolding, transportation and degradation under normal or abnormal environmental conditions (Clare & Saibil, 2013; King & MacRae, 2015).…”

Heat Shock Protein 70 Family in Response to Multiple Abiotic Stresses in the Silkworm

“…Previous studies suggested that HSP70/HSC70 proteins have some characteristic sequences or motifs [42]. To understand the conservation of the characteristic sequences, all the HSP70/HSC70 proteins of B. mori, B. tabaci, and D. melanogaster were aligned by Clus-talX v1.83 [37].…”

“…To understand the conservation of the characteristic sequences, all the HSP70/HSC70 proteins of B. mori, B. tabaci, and D. melanogaster were aligned by Clus-talX v1.83 [37]. The consensus sequences [42] were used to locate the signatures and motifs, and the characteristic alignments were presented in Figure 2. Three signature sequences ('GIDLGTTYS', 'IFDLGGGTFDVSIL,' and 'VGGSTRIPKVQ') were found (Figure 2).…”

A heat shock 70kDa protein MaltHSP70-2 contributes to thermal resistance in Monochamus alternatus (Coleoptera: Cerambycidae): quantification, localization, and functional analysis

“…Our previous investigations identified a suit of HSP genes in M. alternatus larvae induced by a short-term heat shock treatment using comparative transcriptome analysis [ 29 ]. HSP70 subfamily, as principal member of these induced HSP genes, was further characterized in different tissues of M. alternatus larvae [ 28 ]. Among six HSP70 genes of M. alternatus , the increased transcripts of MaltHSP70-2 were at the highest levels upon heat stress, 7109-fold higher than the control levels [ 28 ].…”