猴源葡萄糖调节蛋白78生物信息学分析及其真核表达

doi:10.16431/j.cnki.1671-7236.2021.04.003

Abstract

Abstract: The present study was aimed to explore the physicochemical properties and structural characteristics of chlorocebus sabaeus glucose regulatory protein 78 (GRP78) gene,and to elucidate the molecular chaperone effect and regulatory mechanism of GRP78 gene in porcine epidemic diarrhea virus replication.GRP78 gene was amplified by RT-PCR and inserted into pMD20-T Simple vector for cloning and sequencing,and its amino acids sequence,transmembrane structure,glycosylation site,phosphorylation site,and tertiary structure were predicted and analyzed by bioinformatics methods.The GRP78 gene was inserted into pCDNA 3.1,and transfected into Vero-E6 cells.Western blot was employed to confirm the expression of GRP78 in Vero-E6 cells.Bioinformatics analysis results showed that the total length of GRP78 gene was 1 965 bp,encoding 654 amino acids.The molecular weight of GRP78 protein was 72.33 ku.The theoretical isoeletric point was 5.07,and the molecular formula was C₃₁₈₉H₅₁₅₃N₈₆₅O₁₀₁₉S₁₃.The homology of amino acid sequence of GRP78 gene in Chlorocebus sabaeus with Camelus bactrianus, Canis lupus, Felis catus, Gorilla, Myotis lucifugus, Papio anubis, Sus scrofa, Equus caballus were 99.5% to 99.8%.Genetic analysis showed that gorillas and baboons were the closest relatives.The prediction results of transmembrane region and signal peptide showed that the protein had signal peptide but no transmembrane region.The protein had 6 O-glycosylation sites and 28 phosphorylation sites but no N-glycosylation modification sites,which suggested that GRP78 might have binding sites for PKC and PKA specific protein kinases related to kinase phosphorylation,and might be involved in hexose metabolism and monosaccharide metabolism.

Key words: GRP78 gene; bioinformatics analysis; eukaryotic expression

CLC Number:

Q786

JIAO Wenqiang, LIU Yunchao, XING Guangxu, CHEN Pengju, XU Yindi, WANG Zhifang, ZHANG Qingxian, XU Feng. Bioinformatics Analysis and Eukaryotic Expression of Chlorocebus sabaeus Glucose Regulatory Protein 78[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1170-1178.

References

[1] HETZ C.The unfolded protein response:Controlling cell fate decisions under ER stress and beyond[J].Nature Reviews Molecular Cell Biology,2012,13(2):89-102.
[2] WALTER P,RON D.The unfolded protein response:From stress pathway to homeostatic regulation[J].Science,2011,334(6059):1081-1086.
[3] 樊欣,彭仁.热休克蛋白70:生物学功能与作用机制研究进展[J].生命科学,2019,31(3):270-278. FAN X,PENG R.Heat shock protein 70:Biological function and the mechanism for its function[J].Chinese Bulletin of Life Sciences,2019,31(3):270-278.(in Chinese).
[4] SENFT D,RONAI Z.UPR,autophagy,and mitochondria crosstalk underlines the ER stress response[J].Trends Biochemical Sciences,2015,40(3):141-148.
[5] JUNG K,SAIF L J.Porcine epidemic diarrhea virus infection:Etiology,epidemiology,pathogenesis and immunoprophylaxis[J].Veterinary Journal,2015,204(2):134-143.
[6] WOOD E.An apparently new syndrome of porcine epidemic diarrhea[J].The Veterinary Record,1977,100(12):243-244.
[7] PIJPERS A,VAN NIEUWSTADT A,TERPSTRA C,et al.Porcine epidemic diarrhea virus as a cause of persistent diarrhea in a herd of breeding and finishing pigs[J].The Veterinary Record,1993,132(6):129-131.
[8] NAGY B,NAGY G,MEDER M,et al.Enteroto-xigenic Escherichia coli,rotavirus,porcine epidemic diarrhea virus,adenovirus and calici-like virus in porcine postweaning diarrhea in Hungary[J].Acta Veterinaria Hungarica,1996,44(1):9-19.
[9] TAKAHASHI K,OKADA K,OHSHIMA K,et al.An outbreak of swine diarrhea of a new-type associated with coronavirus-like particles in Japan[J].The Japanese Journal of Veterinary Science,1983,45(6):829-32.
[10] CHAE C,KIM O,CHOI C,et al.Prevalence of porcine epidemic diarrhea virus and transmissible gastroenteritis virus infection in Korean pigs[J].The Veterinary Record,2000,147(21):606-608.
[11] LEE S,LEE C.Outbreak-related porcine epidemic diarrhea virus strains similar to US strains,South Korea[J].Emerging Infectious Diseases,2014,20(7):1223-1226.
[12] MOLE B.Deadly pigs virus slips through US borders[J].Nature,2013,499(7459):388.
[13] SVETLANA P E,BONG S K,BU Y C,et al.Epigallocatechin gallate overcomes resistance to etoposide-induced cell death by targeting the molecular chaperone glucose-regulated protein[J].Cancer Research,2006,66(18):9260-9269.
[14] SCOTT A H,HONG Y R,VICTORIA J M,et al.Quality control autophagy degrade soluble ERAD-resistant conformers of the misfolded membrane protein GnRHR[J].Molecular Cell,2014,54(1):166-179.
[15] SANO R,REED J C.ER stress-induced cell death mechanisms[J].Biochimica et Biophysica Acta,2013,1833(12):3460-3470.
[16] LUO B,LEE A S.The critical roles of endoplasmic reticulum chaperones and unfolded protein response in tumorigenesis and anticancer therapies[J].Oncogene,2013,32(7):805-818.
[17] RUBIOLO J A,LOPEZ-ALONSO H,MARTINEZ P,et al.Yessotoxin induces ER-stress followed by autophagic cell death in Glioma cells mediated by mTOR and BNIP3[J].Cellular Signalling,2014,26(2):419-432.
[18] TOMAR D,PRAJAPATI P,SRIPADA L,et al.TRIM13 regulates Caspase-8 ubiquitination,translocation to autophagosomes and activation durding ER stress induced cell death[J].Biochimica et Biophysica Acta Molecular Cell Research,2013,1833(12):3134-3144.
[19] BECK D,NIESSNER H,SMALLEY K,et al.Vemurafenib potently induces endoplasmic reticulum stress-mediated apoptosis in BRAFV600E melanoma cells[J].Science Signaling,2013,6(260):ra7.
[20] DASH S,CHAVA S,AYDIN Y,et al.Hepatitis C virus infection induces autophagy as a prosurvival mechanism to alleviate hepatic ER-stress response[J].Viruses,2016,8(5):150-178.
[21] SHARMA M,BHATTACHARYYA S,SHARMA K B,et al.Japanese encephalitis virus activates autophagy through XBP1 and ATF6 ER stress sensors in neuronal cells[J].Journal of General Virology,2017,98(5):1027-1039.
[22] LEE Y R,KUO S H,LIN C Y,et al.Dengue virus-induced ER stress is required for autophagy activation,viral replication,and pathogenesis both in vitro and in vivo[J].Scientific Reports,2018,8(1):489-493.
[23] AMBROSE R L,MACKENZIE J M.West Nile virus differentially modulates the unfolded protein response to facilitate replication and immune evasion[J].Journal of Virology,2011,85(6):2723-2732.
[24] ZHOU Y S,GU Y X,QI B Z,et al.Porcine circovirus type 2 capsid protein induces unfolded protein response with subsequent activation of apoptosis[J].Journal Zhejiang University Science B,2017,18(4):316-323.

Bioinformatics Analysis and Eukaryotic Expression of Chlorocebus sabaeus Glucose Regulatory Protein 78

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