猪ADAR2基因全长cDNA克隆、序列特征及表达模式分析

doi:10.11843/j.issn.0366-6964.2021.03.006

Abstract

Abstract: This study aimed to clone the full-length cDNA of the porcine ADAR2 gene and explore its expression pattern in different tissues of pigs. Rapid-amplification of cDNA ends (RACE) was used to clone the full-length cDNA sequence of the ADAR2 gene in Large White pigs and the sequence was analyzed by bioinformatics. Real-time PCR was used to detect the ADAR2 mRNA expression in the heart, liver, lung, kidney, spleen, brain, small intestine, muscle and backfat of 35-day-old pigs. Porcine ADAR2 cDNA sequence of 6 305 bp was cloned, which contained 12 exons and encoded 704 amino acids. The nucleic acid and amino acid sequences shared a high identity (>84%) with that of other mammals including human, chimpanzee, macaque, gibbon, cow, goat and sheep. The deduced ADAR2 had two double-stranded RNA binding motifs and an adenosine deaminase domain. Real-time PCR results showed that the ADAR2 expressed in all the detected tissues, and had the highest expression in the lung. The full-length cDNA sequence of porcine ADAR2 gene was successfully cloned, and it was widely expressed in tissues of pigs. These findings provide a solid foundation for further function study of ADAR2.

Key words: ADAR2, expression, cDNA clone, RACE, swine

CLC Number:

S828.2

ZHANG Yuebo, WANG Ligang, HOU Xinhua, LIU Xin, YAN Hua, ZHANG Longchao, WANG Lixian. The Full-length cDNA Cloning, Sequence Characterization and Expression Pattern Analysis of Porcine ADAR2 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 620-629.

References 41

[1]	张跃博,颜华,王立刚,等.哺乳动物RNA编辑及其检测方法[J].畜牧兽医学报,2018,49(11):2299-2309.ZHANG Y B,YAN H,WANG L G,et al.RNA editing and its detection methods in mammalian[J].Acta Veterinaria et Zootechnica Sinica,2018,49(11):2299-2309.(in Chinese)
[2]	ZHANG Y B,ZHANG L C,YUE J W,et al.Genome-wide identification of RNA editing in seven porcine tissues by matched DNA and RNA high-throughput sequencing[J].J Anim Sci Biotechnol,2019,10:24.
[3]	CHEN J Y,PENG Z Y,ZHANG R L,et al.RNA editome in rhesus macaque shaped by purifying selection[J].PLoS Genet,2014, 10(4):e1004274.
[4]	OUYANG Z Y,REN C,LIU F,et al.The landscape of the A-to-I RNA editome from 462 human genomes[J].Sci Rep,2018, 8(1):12069.
[5]	ZHOU R,YAO W Y,XIE C D,et al.Developmental stage-specific A-to-I editing pattern in the postnatal pineal gland of pigs (Sus scrofa)[J].J Anim Sci Biotechnol,2020,11(1):90.
[6]	WANG L G,LI J N,HOU X H,et al.Genome-wide identification of RNA editing sites affecting intra-muscular fat in pigs[J].Animals, 2020,10(9):1616.
[7]	EISENBERG E,LEVANON E Y.A-to-I RNA editing-immune protector and transcriptome diversifier[J].Nat Rev Genet,2018, 19(8):473-490.
[8]	GATSIOU A,VLACHOGIANNIS N,LUNELLA F F,et al.Adenosine-to-inosine RNA editing in health and disease[J].Antioxid Redox Signal,2018,29(9):846-863.
[9]	FRITZELL K,XU L D,LAGERGREN J,et al.ADARs and editing:The role of A-to-I RNA modi-fication in cancer progression[J]. Semin Cell Dev Biol,2018,79:123-130.
[10]	SONG Y Y,AN O,REN X,et al.RNA editing mediates the functional switch of COPA in a novel mechanism of hepatocarcinogenesis[J/OL].J Hepatol,2020,doi:10.1016/j.jhep.2020.07.021.
[11]	KLAUE Y,KÄLLMAN A M,BONIN M,et al.Biochemical analysis and scanning force microscopy reveal productive and nonproductive ADAR2 binding to RNA substrates[J].RNA,2003,9(7):839-846.
[12]	HIGUCHI M,SINGLE F N,KÖHLER M,et al.RNA editing of AMPA receptor subunit GluR-B:a base-paired intron-exon structure determines position and efficiency[J].Cell,1993,75(7):1361-1370.
[13]	HIGUCHI M,MAAS S,SINGLE F N,et al.Point mutation in an AMPA receptor gene rescues lethality in mice deficient in the RNA-editing enzyme ADAR2[J].Nature,2000,406(6791):78-81.
[14]	HORSCH M,SEEBURG P H,ADLER T,et al.Requirement of the RNA-editing enzyme ADAR2 for normal physiology in mice[J].J Biol Chem,2011,286(21):18614-18622.
[15]	TAN M H,LI Q,SHANMUGAM R,et al.Dynamic landscape and regulation of RNA editing in mammals[J].Nature, 2017, 550(7675):249-254.
[16]	SINGH M,KESTERSON R A,JACOBS M M,et al.Hyperphagia-mediated obesity in transgenic mice mis-expressing the RNA-editing enzyme ADAR2[J].J Biol Chem,2007,282(31):22448-22459.
[17]	CHEN C J,CHEN H,ZHANG Y,et al.TBtools:an integrative toolkit developed for interactive analyses of big biological data[J].Molecular Plant,2020,13(8):1194-1202.
[18]	PICARDI E,D'ERCHIA A M,GIUDICE C L,et al.REDIportal:a comprehensive database of A-to-I RNA editing events in humans[J].Nucleic Acids Res,2017,45(D1):D750-D757.
[19]	OAKES E,ANDERSON A,COHEN-GADOL A,et al.Adenosine deaminase that acts on RNA 3(ADAR3) binding to glutamate receptor subunit B pre-mRNA inhibits RNA editing in glioblastoma[J].J Biol Chem,2017,292(10):4326-4335.
[20]	SLAVOV D,GARDINER K.Phylogenetic com-parison of the pre-mRNA adenosine deaminase ADAR2 genes and transcripts:conservation and diversity in editing site sequence and alternative splicing patterns[J].Gene,2002,299(1-2):83-94.
[21]	BURSET M,SELEDTSOV I A,SOLOVYEV V V.Analysis of canonical and non-canonical splice sites in mammalian genomes[J]. Nucleic Acids Res,2000,28(21):4364-4375.
[22]	BLOW M,FUTREAL P A,WOOSTER R,et al.A survey of RNA editing in human brain[J].Genome Res,2004,14(12):2379-2387.
[23]	SANSAM C L,WELLS K S,EMESON R B.Modulation of RNA editing by functional nucleolar sequestration of ADAR2[J].Proc Natl Acad Sci U S A,2003,100(24):14018-14023.
[24]	BEHM M,WAHLSTEDT H,WIDMARK A,et al.Accumulation of nuclear ADAR2 regulates adenosine-to-inosine RNA editing during neuronal development[J].J Cell Sci,2017,130(4):745-753.
[25]	BLOM N,SICHERITZ-PONTÉN T,GUPTA R,et al.Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence[J].Proteomics,2004,4(6):1633-1649.
[26]	MELCHER T,MAAS S,HERB A,et al.A mammalian RNA editing enzyme[J].Nature,1996,379(6564):460-464.
[27]	张耀丹,汪伟,李继红,等.鸡源ADAR2基因克隆及其抗新城疫病毒作用研究[J].中国动物传染病学报,2017,25(2):35-42.ZHANG Y D,WANG W,LI J H,et al.Identification of chicken adenosine acting on RNA (ADAR2) and its anti-viral effect on newcastle disease virus[J].Chinese Journal of Animal Infectious Diseases,2017,25(2):35-42.(in Chinese)
[28]	YU Z,CHEN T Y,CAO X T.RNA editing by ADAR1 marks dsRNA as "self"[J].Cell Res,2015,25(12):1283-1284.
[29]	YANAI M,KOJIMA S,SAKAI M,et al.ADAR2 is involved in self and nonself recognition of borna disease virus genomic RNA in the nucleus[J].J Virol,2020,94(6):e01513-19.
[30]	NAKAHAMA T,KATO Y,KIM J I,et al.ADAR1-mediated RNA editing is required for thymic self-tolerance and inhibition of autoimmunity[J].EMBO Rep,2018,19(12):e46303.
[31]	张跃博,欧阳峰正,王立刚,等.猪ADAR1基因cDNA全长克隆、序列信息及组织表达分析[J].畜牧兽医学报,2019,50(06):1135-1144.ZHANG Y B,OUYANG F Z,WANG L G,et al.The full-length cloning,sequence information and expression analysis of porcine ADAR1 gene[J].Acta Veterinaria et Zootechnica Sinica,2019,50(6):1135-1144.(in Chinese)
[32]	GALLO A,LOCATELLI F.ADARs:allies or enemies? The importance of A-to-I RNA editing in human disease:from cancer to HIV-1[J].Biol Rev Camb Philos Soc,2012,87(1):95-110.
[33]	GALIPON J,ISHⅡ R,SUZUKI Y,et al.Differential binding of three major human ADAR isoforms to coding and long non-coding transcripts[J].Genes,2017,8(2):68.
[34]	LEVANON E Y,EISENBERG E,YELIN R,et al.Systematic identification of abundant A-to-I editing sites in the human transcriptome[J].Nat Biotechnol,2004,22(8):1001-1005.
[35]	SOMMER B,KÖHLER M,SPRENGEL R,et al.RNA editing in brain controls a determinant of ion flow in glutamate-gated channels[J].Cell,1991,67(1):11-19.
[36]	RUETER S M,DAWSON T R,EMESON R B.Regulation of alternative splicing by RNA editing[J].Nature,1999, 399(6731):75-80.
[37]	TERAJIMA H,YOSHITANE H,OZAKI H,et al.ADARB1 catalyzes circadian A-to-I editing and regulates RNA rhythm[J].Nat Genet,2017,49(1):146-151.
[38]	TERAJIMA H,YOSHITANE H,YOSHIKAWA T,et al.A-to-I RNA editing enzyme ADAR2 regulates light-induced circadian phase-shift[J].Sci Rep,2018,8(1):14848.
[39]	田宇,潘玉琢,杜超,等.胶质瘤中RNA编辑酶ADAR2,ADAR3 mRNA表达及意义[J].中国实验诊断学,2006,10(1):23-26.TIAN Y,PAN Y Z,DU C,et al.The expression of RNA editing deaminase ADAR2,ADAR3 mRNA on human glioma cell lines[J]. Chinese Journal of Laboratory Diagnosis,2006,10(1):23-26.(in Chinese)
[40]	MAAS S,PATT S,SCHREY M,et al.Underediting of glutamate receptor GluR-B mRNA in malignant gliomas[J].Proc Natl Acad Sci U S A,2001,98(25):14687-14692.
[41]	CENCI C,BARZOTTI R,GALEANO F,et al.Down-regulation of RNA editing in pediatric astrocytomas:ADAR2 editing activity inhibits cell migration and proliferation[J].J Biol Chem,2008,283(11):7251-7260.

The Full-length cDNA Cloning, Sequence Characterization and Expression Pattern Analysis of Porcine ADAR2 Gene

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