水貂AANAT基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2023.04.003

Abstract

Abstract: 【Objective】 The purpose of this experiment was to clone and bioinformatically analyze the gene of the rate-limiting enzyme aralkylamine-N-acetyltransferase (AANAT) for N-acetyl-5-methoxytryptamine (MT) synthesis in American mink (Neovison vison),so as to provide a reference for biological functions in mink.【Method】 DNA was extracted from the caudal vein of mink,the sequence of AANAT gene was cloned by homologous recombination,the sequence of CDS region was analyzed,the amino acid sequence of AANAT gene was deduced for similarity alignment and phylogenetic tree construction,and the AANAT protein was analyzed by bioinformatics.【Result】 The sequence of AANAT gene in mink was about 1 631 bp,and the sequence of CDS region was 504 bp,which could encode 167 amino acids.The amino acid sequence similarity of AANAT gene in mink was 98.2% with Mustela putorius furo,and the phylogenetic tree analysis also showed that it was the closest relative to Mustela putorius furo.Bioinformatics analysis results showed that AANAT protein in mink was a hydrophobic protein without transmembrane domain and signal peptide,mainly distributed in the cytoplasm,and had 5 antigen-binding sites as a non-secreted protein.AANAT protein contained multiple post-translational modification sites,such as phosphorylation and glycosylation.AANAT protein contained a conserved domain of the N-acyltransferase superfamily,which was closely related to mammalian circadian rhythms.The secondary structure of AANAT protein was dominated by random coil,and the tertiary structure contained several conserved catalytic residues.Protein network interaction analysis results showed that AANAT protein could interact with multipleserotonin synthesis-related proteins.【Conclusion】 The sequence of AANAT gene in mink was successfully obtained.The structure and specific modification sites of AANAT protein might participate in the regulation of MT biosynthesis by localizing acetyl-CoA and 5-hydroxytryptamine.The reults provided a reference for research on the regulation mechanism of AANAT gene on embryonic diapause and improving fecundity in mink.

Key words: mink; AANAT gene; cloning; bioinformatics analysis; protein structure

CLC Number:

FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.

References

[1] WANG C,LI F,ZHANG Z,et al.Recent research progress in China on Haemonchus contortus[J].Frontiers in Microbiology,2017,8:1509.
[2] 郝丹,苏国生,吴晓平,等.世界水貂遗传育种的研究进展[J].黑龙江畜牧兽医,2021,7:43-47. HAO D,SU G S,WU X P,et al.Progress in research on genetic breeding of worldwide mink[J].Heilongjiang Animal Science and Veterinary Medicine,2021,7:43-47.(in Chinese)
[3] HICKMAN A B,KLEIN D C,DYDA F.Melatonin biosynthesis:The structure of serotonin N-acetyltransferase at 2.5 A resolution suggests a catalytic mechanism[J].Molecular Cell,1999,3(1):23-32.
[4] SZEWCZYK-GOLEC K,WOZNIAK A,REITER R J.Inter-relationships of the chronobiotic,melatonin,with leptin and adiponectin:Implications for obesity[J].Journal of Pineal Research,2015,59(3):277-291.
[5] KAMRUZZAMAN A S M,HIRAGAKI S,WATARI Y,et al.Clock-controlled arylalkylamine N-acetyltransferase (aaNAT) regulates circadian rhythms of locomotor activity in the American cockroach,Periplaneta americana,via melatonin/MT2-like receptor[J].Journal of Pineal Research,2021,71(2):e12751.
[6] KLEIN D C.Photoneural regulation of the mammalian pineal gland[J].Ciba Foundation Symposium,1985,117:38-56.
[7] MARONDE E,SCHOMERUS C,STEHLE J H,et al.Control of CREB phosphorylation and its role for induction of melatonin synthesis in rat pinealocytes[J].Biology of the Cell,1997,89(8):505-511.
[8] HO A K,CHIK C L.Modulation of aanat gene transcription in the rat pineal gland[J].Journal of Neurochemistry,2010,112(2):321-331.
[9] FAILLACE M P,CUTRERA R,SARMIENTO M I,et al.Evidence for local synthesis of melatonin in golden hamster retina[J].Neuroreport,1995,6(15):2093-2095.
[10] TAGLIAFERRI V,ROMUALDI D,SCARINCI E,et al.Melatonin treatment may be able to restore menstrual cyclicity in women with PCOS:A pilot study[J].Reproductive Sciences,2018,25(2):269-275.
[11] ZHANG W L,ZHANG Z L,PENG J,et al.Expression of arylalkylamine N-acetyltransferase (AANAT) and acetylserotonin O-methyltransferase (ASMT) in the corpus luteum of pregnant sows and synthesis of melatonin in luteal cells[J].Cell and Tissue Research,2022,388(1):167-179.
[12] RENFREE M B,SHAW G.Diapause[J].Annual Review of Physiology,2000,62:353-375.
[13] 韩玉萍,赵向远,范冰峰,等.基于高通量测序的水貂胚胎滞育期和激活期卵巢转录组分析[J].中国畜牧兽医,2021,48(11):4035-4046. HAN Y P,ZHAO X Y,FAN B F,et al.Transcriptomic analysis of mink ovaries at embryonic diapause and activation stages based on high-throughput sequence technology[J].China Animal Husbandry & Veterinary Medicine,2021,48(11):4035-4046.(in Chinese)
[14] 韩玉萍,李文,范冰峰,等.胚胎滞育的研究进展[J].特产研究,2020,42(6):60-67. HAN Y P,LI W,FAN B F,et al.Research progress on embryo diapause[J].Special Wild Economic Animal and Plant Research, 2020,42(6):60-67.(in Chinese)
[15] TAKEDA M,SUZUKI T.Circadian and neuroendocrine basis of photoperiodism controlling diapause in insects and mites:A review[J].Frontiers in Physiology,2022,13:867621.
[16] CARLOMAGNO G,MININI M,TILOTTA M,et al.From implantation to birth:Insight into molecular melatonin functions[J].International Journal of Molecular Sciences,2018,19(9):2802.
[17] DENG L,LI C,CHEN L,et al.Research advances on embryonic diapause in mammals[J].Animal Reproduction Science,2018,198:1-10.
[18] TAN D X,MANCHESTER L C,QIN L,et al.Melatonin:A mitochondrial targeting molecule involving mitochondrial protection and dynamics[J].International Journal of Molecular Sciences,2016,17(12):2124.
[19] KLEIN D C,COON S L,ROSEBOOM P H,et al.The melatonin rhythm-generating enzyme:Molecular regulation of serotonin N-acetyltransferase in the pineal gland[J].Recent Progress in Hormone Research,1997,52:307-357.
[20] VETTING M W,DE CARVALHO L P S,YU M,et al.Structure and functions of the GNAT superfamily of acetyltransferases[J].Archives of Biochemistry and Biophysics,2005,433(1):212-226.
[21] VETTING M W,RODERICK S L,YU M,et al.Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of N-acetyltransferases[J].Protein Science:A Publication of the Protein Society,2003,12(9):1954-1959.
[22] VETTING M W,HEGDE S S,JAVID-MAJD F,et al.Aminoglycoside 2'-N-acetyltransferase from Mycobacterium tuberculosis in complex with coenzyme A and aminoglycoside substrates[J].Nature Structural Biology,2002,9(9):653-658.
[23] SUGDEN D,VANECEK J,KLEIN D C,et al.Activation of protein kinase C potentiates isoprenaline-induced cyclic AMP accumulation in rat pinealocytes[J].Nature,1985,314(6009):359-361.
[24] COON S L,ROSEBOOM P H,BALER R,et al.Pineal serotonin N-acetyltransferase:Expression cloning and molecular analysis[J].Science,1995,270(5242):1681-1683.
[25] POIREL V J,BOGGIO V,DARDENTE H,et al.Contrary to other non-photic cues,acute melatonin injection does not induce immediate changes of clock gene mRNA expression in the rat suprachiasmatic nuclei[J].Neuroscience,2003,120(3):745-755.
[26] EICHLER J.Protein glycosylation[J].Current Biology,2019,29(7):R229-R231.
[27] SCHJOLDAGER K T,NARIMATSU Y,JOSHI H J,et al.Global view of human protein glycosylation pathways and functions[J].Nature Reviews Molecular Cell Biology,2020,21(12):729-749.
[28] MACLEA K S,KRIESER R J,EASTMAN A.Structural requirements of human DNase Ⅱ alpha for formation of the active enzyme:The role of the signal peptide,N-glycosylation,and disulphide bridging[J].Biochemical Journal,2003,371:867-876.
[29] WANG B,DAI T,SUN W,et al.Protein N-myristoylation:Functions and mechanisms in control of innate immunity[J].Cellular and Molecular Immunology,2021,18(4):878-888.
[30] FARAZI T A,WAKSMAN G,GORDON J I.The biology and enzymology of protein N-myristoylation[J].The Journal of Biological Chemistry,2001,276(43):39501-39504.
[31] 杨爱玲,李广栋,吴昊,等.褪黑素合成酶AANAT/ASMT基因过表达绵羊生物安全评价研究[J].畜牧兽医学报,2020,51(7):1563-1572. YANG A L,LI G D,WU H,et al.Study of biosafety evaluation melatonin AANAT/ASMT overexpressed sheep[J].Acta Veterinaria et Zootechnica Sinica,2020,51(7):1563-1572.(in Chinese)
[32] GUENTER J,LENARTOWSKI R.Molecular characteristic and physiological role of DOPA-decarboxylase[J].Postepy Higieny I Medycyny Doswiadczalnej, 2016,70:1424-1440.
[33] SON S Y,MA J,KONDOU Y,et al.Structure of human monoamine oxidase A at 2.2-A resolution:The control of opening the entry for substrates/inhibitors[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(15):5739-5744.
[34] THOMPSON M A,MOON E,KIM U J,et al.Human indolethylamine N-methyltransferase:cDNA cloning and expression,gene cloning,and chromosomal localization[J].Genomics,1999,61(3):285-297.

Cloning and Bioinformatics Analysis of AANAT Gene in Mink

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