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

Abstract

BackgroundThe cold shock domain (CSD) containing proteins (CSDPs) are one group of the evolutionarily conserved nucleic acid-binding proteins widely distributed in bacteria, plants, animals, and involved in various cellular processes, including adaptation to low temperature, cellular growth, nutrient stress and stationary phase.MethodologyThe cDNA of a novel CSDP was cloned from Zhikong scallop Chlamys farreri (designated as CfCSP) by expressed sequence tag (EST) analysis and rapid amplification of cDNA ends (RACE) approach. The full length cDNA of CfCSP was of 1735 bp containing a 927 bp open reading frame which encoded an N-terminal CSD with conserved nucleic acids binding motif and a C-terminal domain with four Arg-Gly-Gly (RGG) repeats. The CSD of CfCSP shared high homology with the CSDs from other CSDPs in vertebrate, invertebrate and bacteria. The mRNA transcripts of CfCSP were mainly detected in the tissue of adductor and also marginally detectable in gill, hepatopancreas, hemocytes, kidney, mantle and gonad of healthy scallop. The relative expression level of CfCSP was up-regulated significantly in adductor and hemocytes at 1 h and 24 h respectively after low temperature treatment (P<0.05). The recombinant CfCSP protein (rCfCSP) could bind ssDNA and in vitro transcribed mRNA, but it could not bind dsDNA. BX04, a cold sensitive Escherichia coli CSP quadruple-deletion mutant, was used to examine the cold adaptation ability of CfCSP. After incubation at 17°C for 120 h, the strain of BX04 containing the vector pINIII showed growth defect and failed to form colonies, while strain containing pINIII-CSPA or pINIII-CfCSP grew vigorously, indicating that CfCSP shared a similar function with E. coli CSPs for the cold adaptation.ConclusionsThese results suggest that CfCSP is a novel eukaryotic cold-regulated nucleic acid-binding protein and may function as an RNA chaperone in vivo during the cold adaptation process.

Transcriptomic and proteomic analyses on the supercooling ability and mining of antifreeze proteins of the Chinese white wax scale insect

“…To prevent freezing during winter, E. pela must undergo extensive dehydration and must clear ice nuclei substances from the gut and hemolymph (Carrasco et al ., ). Cold adaptation proteins have also been implicated in tolerance to low temperatures (De Leeuw et al ., ; Yang et al ., ). In the present study, these genes were identified in the transcriptome but were absent in the proteome.…”

Proteomic analysis of early salt stress responsive proteins in alfalfa roots and shoots

“…Two nucleic acid binding proteins (NABP1, spots 15, R35) showed altered abundance in both the shoots and roots under salt stress. NABP is a small and highly conserved protein with nucleic acid chaperone activity that binds single-stranded nucleic acids [ 69 ]. One group of NABPs is the cold shock domain (CSD) containing proteins, and these CSDPs are involved in various cellular processes, including adaptation to low temperatures, cellular growth, nutrient stress, and the stationary phase [ 70 , 71 ].…”

The Polymorphism in the Promoter of HSP70 Gene Is Associated with Heat Tolerance of Two Congener Endemic Bay Scallops (Argopecten irradians irradians and A. i. concentricus)

“…The SYBR Green RT-PCR assay was carried out and analyzed as previously described [23] . The comparative average cycle threshold method was used to analyze the mRNA expression level of AiHSP70, and the value stood for an n-fold difference relative to the calibrator [24] .…”