The Gln32Lys Polymorphism in HSP22 of Zhikong Scallop Chlamys farreri Is Associated with Heat Tolerance

Abstract

BackgroundHeat shock protein 22 is a member of small heat shock proteins with molecular chaperone activity. Though their multiple functions have been well characterized, there is no report about the association between the polymorphisms of HSP22 and heat tolerance.MethodologyThree single nucleotide polymorphisms were identified in HSP22 from scallop Chlamys farreri (CfHSP22), and the +94 C-A locus was found to be nonsynonymous. Three genotypes at locus +94, A/A, A/C and C/C, were revealed by using Bi-PASA PCR analysis, and their frequencies were 19.5%, 27.6% and 52.9% in the heat resistant stock, while 9.3%, 17.4% and 73.3% in the heat susceptible stock, respectively. The frequency differences of the three genotypes were significant (P<0.05) between the two stocks. After incubating at 30°C for 84 h, the cumulative mortality of scallops with +94 C/C genotype and +94 A/C genotypes was 95% and 90%, respectively, which was significantly higher (P<0.01) than that of scallops with +94 A/A genotype (70%). The molecular chaperone activity of two His-tagged fusion proteins, rCfHSP22Q with +94 C/C genotype and rCfHSP22K with +94 A/A genotype were analyzed by testing the ability of protecting citrate synthase (CS) against thermal inactivation in vitro. After incubated with rCfHSP22Q or rCfHSP22K at 38°C for 1 h, the activity of CS lost 50% and 45%, and then recovered to 89% and 95% of the original activity following 1 h restoration at 22°C, respectively, indicating that the mutation from Gln to Lys at this site might have an impact on molecular chaperone activities of CfHSP22.ConclusionsThese results implied that the polymorphism at locus +94 of CfHSP22 was associated with heat tolerance of scallop, and the +94 A/A genotype could be a potential marker available in future selection of Zhikong scallop with heat tolerance.

Sequencing‐based gene network analysis provides a core set of gene resource for understanding thermal adaptation in Zhikong scallop Chlamys farreri

“…Its aquaculture industry has suffered substantial losses because of heavy summer mortalities over the past few decades (Yang et al . ). Several studies have been conducted in an attempt to understand the molecular responses of C. farreri to heat stress (Qu et al .…”

“…; Yang et al . ). However, these studies have been limited to specific genes that have been well studied in model organisms.…”

A Novel Cold-Regulated Cold Shock Domain Containing Protein from Scallop Chlamys farreri with Nucleic Acid-Binding Activity

“…The recombinant plasmid (pET-30a-CfCSP) was transformed into E. coli BL21 (DE3)-pLysS (Novagen). The recombinant CfCSP protein was purified with a Ni 2+ chelating Sepharose column, and the purified protein was refolded as previously described [45] . Then the resultant protein was separated by reducing 15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and visualized with Coomassie Bright Blue R250.…”

The immune system and its modulation mechanism in scallop

“…Some polymorphisms of HSPs have been found to be significantly associated with heat tolerance. For example, the polymorphism at locus þ94 of CfHSP22 gene was associated with heat tolerance of C. farreri [95]. And the SNPs in AiHSP70 and AiHSP90 promoter were associated with thermal adaptation of different congener endemic A. irradians [96,97].…”