牦牛StAR基因克隆及其生物信息学与组织表达特性的研究

doi:10.11843/j.issn.0366-6964.2021.09.008

Abstract

Abstract: The objective of this study was to obtain the coding sequence (CDS) of yak StAR gene, conduct bioinformatics analysis and investigate its mRNA expression characteristics in different tissues. Yak samples were collected from local slaughter house, including heart, liver, spleen, lung, kidney, ovary, oviduct and uterus of adult female yaks (n=5), ovary of yaks at different ages (fetal, 1-year-old and 2-years-old) (n=3), ovary of yaks during different periods of estrous cycles (follicular phase and luteal phase) (n=3), the luteal phase ovary of cattle (n=3). Granulosa cells of yaks were cryopreserved previously. StAR gene was cloned using yak ovarian cDNA as template during luteal phase by reverse transcriptase-polymerase chain reaction (PCR) and its bioinformatics properties were analyzed by bioinformatics software such as MEGA7.0 and ExPASy-ProtParam etc. The tissue expression characteristics of yak StAR gene were detected by quantitative real-time PCR. The results showed that the CDS of yak StAR gene was 858 bp, encoding 285 amino acids. StAR protein was positively charged and belonged to alkaline hydrophilic stable protein, which had no transmembrane structure and signal peptide. It mainly existed in cytoplasm and mitochondria. StAR gene was highly conserved between different species, which accorded with the law of species evolution. The expression level of yak StAR gene was the highest in ovary (P<0.01). The expression level of StAR gene in the ovary of 2-years-old was significantly higher than that of fetal and 1-year-old (P<0.01). Its expression in yak ovary during luteal phase was significantly higher than that during follicular phase (P<0.01), and was significantly higher in cattle ovary than in yak ovary during luteal phase(P<0.01). During the in vitro culture (IVC) of granulosa cells, the expression of StAR gene increased gradually and reached the peak level at 24 h IVC (P<0.01), and then significantly decreased. The results showed that StAR gene was relatively conservative in the course of animal evolution. Its expression level was greatest in the ovary of yak, and changing with age- and estrous cycle-specific characteristics, which indicate that StAR gene may be involved in the regulation of yak reproduction associated with ovary and corpus luteum function.

Key words: yak, StAR gene, gene cloning, tissue expression, ovary

CLC Number:

S823.85

LIU Yu, WU Jianfei, LI Heng, JING Tian, LU Jianyuan, ZI Xiangdong. Cloning and Analysis of Bioinformatics and Tissue Expression Characteristics of Yak StAR Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2452-2463.

References 49

[1]	CLARK B J, WELLS J, KING S R, et al.The purification, cloning, and expression of a novel luteinizing hormone-induced mitochondrial protein in MA-10 mouse Leydig tumor cells:characterization of the steroidogenic acute regulatory protein (StAR)[J].J Biol Chem, 1994, 269(45):28314-28322.
[2]	TUCKEY R C, HEADLAM M J, BOSE H S, et al.Transfer of cholesterol between phospholipid vesicles mediated by the steroidogenic acute regulatory protein (StAR)[J].J Biol Chem, 2002, 277(49):47123-47128.
[3]	MILLER W L, BOSE H S.Early steps in steroidogenesis:intracellular cholesterol trafficking[J].J Lipid Res, 2011, 52(12):2111-2135.
[4]	LIN D, SUGAWARA T, STRAUSS III J F, et al.Role of steroidogenic acute regulatory protein in adrenal and gonadal steroidogenesis[J].Science, 1995, 267(5205):1828-1831.
[5]	KANG E, KIM Y M, KIM G H, et al.Mutation spectrum of STAR and the founder effect of p.Q258* in Korean patients with congenital lipoid adrenal hyperplasia[J].Mol Med, 2017, 23:149-154.
[6]	LEERS-SUCHETA S, STOCCO D M, AZHAR S.Down-regulation of steroidogenic acute regulatory (StAR) protein in rat Leydig cells:implications for regulation of testosterone production during aging[J].Mech Ageing Dev, 1999, 107(2):197-203.
[7]	STOCCO D M.StAR protein and the regulation of steroid hormone biosynthesis[J].Annu Rev Physiol, 2001, 63:193-213.
[8]	HASEGAWA T, ZHAO L P, CARON K M, et al.Developmental roles of the steroidogenic acute regulatory protein (StAR) as revealed by StAR knockout mice[J].Mol Endocrinol, 2000, 14(9):1462-1471.
[9]	WELLER M M D C A, FORTES M R S, MARCONDES M I, et al.Effect of maternal nutrition and days of gestation on pituitary gland and gonadal gene expression in cattle[J].J Dairy Sci, 2016, 99(4):3056-3071.
[10]	杨沛方, 周志楠, 敖叶, 等.黔北麻羊StAR基因的克隆、生物信息学分析及在单、多羔黔北麻羊不同性腺组织中的表达[J].中国畜牧杂志, 2020, 56(11):78-84.YANG P F, ZHOU Z N, AO Y, et al.Cloning, bioinformatics analysis and expression of StAR gene in Qianbei brown goat[J].Chinese Journal of Animal Science, 2020, 56(11):78-84.(in Chinese)
[11]	ZI X D, LU J Y, ZHOU H, et al.Comparative analysis of ovarian transcriptomes between prolific and non-prolific goat breeds via high-throughput sequencing[J].Reprod Domest Anim, 2018, 53(2):344-351.
[12]	PILON N, DANEAU I, BRISSON C, et al.Porcine and bovine steroidogenic acute regulatory protein (StAR) gene expression during gestation[J].Endocrinology, 1997, 138(3):1085-1091.
[13]	HARTUNG S, RUST W, BALVERS M, et al.Molecular cloning and in vivo expression of the bovine steroidogenic acute regulatory protein[J].Biochem Biophys Res Commun, 1995, 215(2):646-653.
[14]	SHANG G H, PENG X Y, JI C, et al.Steroidogenic acute regulatory protein and luteinizing hormone are required for normal ovarian steroidogenesis and oocyte maturation in zebrafish[J].Biol Reprod, 2019, 101(4):760-770.
[15]	JUENGEL J L, MEBERG B M, TURZILLO A M, et al.Hormonal regulation of messenger ribonucleic acid encoding steroidogenic acute regulatory protein in ovine corpora lutea[J].Endocrinology, 1995, 136(12):5423-5429.
[16]	欧阳宏佳, 孙敬帅, 江丹莉, 等.水禽StAR基因克隆、表达及其对睾丸发育的影响[J].畜牧兽医学报, 2020, 51(12):3013-3022.OUYANG H J, SUN J S, JIANG D L, et al.Cloning and expression of waterfowl StAR gene and its effect on testicular development[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(12):3013-3022.(in Chinese)
[17]	QIU Q, ZHANG G J, MA T, et al.The yak genome and adaptation to life at high altitude[J].Nat Genet, 2012, 44(8):946-949.
[18]	ZI X D.Reproduction in female yaks (Bos grunniens) and opportunities for improvement[J].Theriogenology, 2003, 59(5-6):1303-1312.
[19]	KUMAR S, STECHER G, TAMURA K.MEGA7:molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Mol Biol Evol, 2016, 33(7):1870-1874.
[20]	洪磊, 陈祥, 唐文, 等.黔北麻羊SRD5A2基因干扰载体构建及其对产羔相关基因表达的影响[J].畜牧兽医学报, 2020, 51(12):2980-2990.HONG L, CHEN X, TANG W, et al.Construction of SRD5A2 gene interference vector and its effect on the expression of lambing related genes in Qianbei Ma goats[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(12):2980-2990.(in Chinese)
[21]	MANNA P R, STETSON C L, SLOMINSKI A T, et al.Role of the steroidogenic acute regulatory protein in health and disease[J].Endocrine, 2016, 51(1):7-21.
[22]	MILLER W L.Steroidogenic acute regulatory protein (StAR), a novel mitochondrial cholesterol transporter[J].Biochim Biophys Acta, 2007, 1771(6):663-676.
[23]	CHEN H, ZHANG Q R, CHEN R M, et al.Lipoid congenital adrenal hyperplasia due to steroid acute regulatory protein (STAR) variants in Three Chinese patients[J].J Steroid Biochem Mol Biol, 2020, 200:105635.
[24]	FLEURY A, MATHIEU A P, DUCHARME L, et al.Phosphorylation and function of the hamster adrenal steroidogenic acute regulatory protein (StAR)[J].J Steroid Biochem Mol Biol, 2004, 91(4-5):259-271.
[25]	ARTEMENKO I P, ZHAO D, HALES D B, et al.Mitochondrial processing of newly synthesized steroidogenic acute regulatory protein (StAR), but not total StAR, mediates cholesterol transfer to cytochrome P450 side chain cleavage enzyme in adrenal cells[J].J Biol Chem, 2001, 276(49):46583-46596.
[26]	BOSE H S, LINGAPPA V R, MILLER W L.Rapid regulation of steroidogenesis by mitochondrial protein import[J].Nature, 2002, 417(6884):87-91.
[27]	BAUER M P, BRIDGHAM J T, LANGENAU D M, et al.Conservation of steroidogenic acute regulatory (StAR) protein structure and expression in vertebrates[J].Mol Cell Endocrinol, 2000, 168(1-2):119-125.
[28]	TSUJISHITA Y, HURLEY J H.Structure and lipid transport mechanism of a StAR-related domain[J].Nat Struct Biol, 2000, 7(5):408-414.
[29]	SCHRICK K, BRUNO M, KHOSLA A, et al.Shared functions of plant and mammalian StAR-related lipid transfer (START) domains in modulating transcription factor activity[J].BMC Biol, 2014, 12:70.
[30]	CATERINO M, RUOPPOLO M, MANDOLA A, et al.Protein-protein interaction networks as a new perspective to evaluate distinct functional roles of voltage-dependent anion channel isoforms[J].Mol Biosyst, 2017, 13(12):2466-2476.
[31]	BOSE M, WHITTAL R M, MILLER W L, et al.Steroidogenic activity of StAR requires contact with mitochondrial VDAC1 and phosphate carrier protein[J].J Biol Chem, 2008, 283(14):8837-8845.
[32]	BAYNE R A L, DA SILVA S J M, ANDERSON R A.Increased expression of the FIGLA transcription factor is associated with primordial follicle formation in the human fetal ovary[J].Mol Hum Reprod, 2004, 10(6):373-381.
[33]	WANG Z P, LIU C Y, ZHAO Y G, et al.FIGLA, LHX8 and SOHLH1 transcription factor networks regulate mouse oocyte growth and differentiation[J].Nucleic Acids Res, 2020, 48(7):3525-3541.
[34]	LI H L, XU W T, ZHANG N, et al.Two Figla homologues have disparate functions during sex differentiation in half-smooth tongue sole (Cynoglossus semilaevis)[J].Sci Rep, 2016, 6:28219.
[35]	NA X X, MAO Y, TANG Y M, et al.Identification and functional analysis of fourteen NR5A1 variants in patients with the 46 XY disorders of sex development[J].Gene, 2020, 760:145004.
[36]	SANDHOFF T W, HALES D B, HALES K H, et al.Transcriptional regulation of the rat steroidogenic acute regulatory protein gene by steroidogenic factor 1[J].Endocrinology, 1998, 139(12):4820-4831.
[37]	MUTHUSAMY S, ANDERSSON S, KIM H J, et al.Estrogen receptorβ and 17β-hydroxysteroid dehydrogenase type 6, a growth regulatory pathway that is lost in prostate cancer[J].Proc Natl Acad Sci U S A, 2011, 108(50):20090-20094.
[38]	AL-BADER M, KILARKAJE N.Effects of bleomycin, etoposide and cisplatin treatment on Leydig cell structure and transcription of steroidogenic enzymes in rat testis[J].Eur J Pharmacol, 2015, 747:150-159.
[39]	CLARK B J.ACTH action on StAR biology[J].Front Neurosci, 2016, 10:547.
[40]	SECHMAN A, GRZEGORZEWSKA A K, GRZESIAK M, et al.Nitrophenols suppress steroidogenesis in prehierarchical chicken ovarian follicles by targeting STAR, HSD3B1, and CYP19A1 and downregulating LH and estrogen receptor expression[J].Domest Anim Endocrinol, 2020, 70:106378.
[41]	PRUCHA M S, MARTYNIUK C J, DOPERALSKI N J, et al.Steroidogenic acute regulatory protein transcription is regulated by estrogen receptor signaling in largemouth bass ovary[J].Gen Comp Endocrinol, 2020, 286:113300.
[42]	刘倩, 马晓, 岑双双, 等.中华鳖StAR基因的克隆、表达及多克隆抗体制备[J].水产学报, 2020, 44(11):1813-1823.LIU Q, MA X, CEN S S, et al.Molecular cloning, expression analysis and polyclonal antibody preparation of StAR gene in Chinese soft-shelled turtle (Pelodiscus sinensis)[J].Journal of Fisheries of China, 2020, 44(11):1813-1823.(in Chinese)
[43]	MICHALOVIC L, CURRIN L, GUTIERREZ K, et al.Granulosa cells of prepubertal cattle respond to gonadotropin signaling and upregulate genes that promote follicular growth and prevent cell apoptosis[J].Mol Reprod Dev, 2018, 85(12):909-920.
[44]	JAHROMI M S, TEHRANI F R, NOROOZZADEH M, et al.Elevated expression of steroidogenesis pathway genes;CYP17, GATA6 and StAR in prenatally androgenized rats[J].Gene, 2016, 593(1):167-171.
[45]	PRAKASH B S, SARKAR M, PAUL V, et al.Postpartum endocrinology and prospects for fertility improvement in the lactating riverine buffalo (Bubalus bubalis) and yak (Poephagus grunniens L.)[J].Livest Prod Sci, 2005, 98(1-2):13-23.
[46]	毛宁, 王嘉福, 张福平, 等.香猪卵巢StAR和CYP11A1基因的差异表达研究[J].中国畜牧兽医, 2018, 45(5):1137-1144.MAO N, WANG J F, ZHANG F P, et al.Study on differential expression of StAR and CYP11A1 genes in Xiang Pig ovary[J].China Animal Husbandry & Veterinary Medicine, 2018, 45(5):1137-1144.(in Chinese)
[47]	兰道亮, 熊显荣, 陈亚冰, 等.牦牛体外成熟卵母细胞差异转录组学研究[J].中国科学:生命科学, 2017, 47(10):1099-1112.LAN D L, XIONG X R, CHEN Y B, et al.Differential transcriptome analysis of yak oocytes in vitro maturation[J].Scientia Sinica Vitae, 2017, 47(10):1099-1112.(in Chinese)
[48]	何翃闳, 潘阳阳, 张慧珠, 等.FSH对牦牛卵母细胞EGF、EGFR表达及其细胞凋亡的影响[J].畜牧兽医学报, 2018, 49(9):1899-1907.HE H H, PAN Y Y, ZHANG H Z, et al.The effects of follicle-stimulating hormone (FSH) on the expression of EGF and EGFR in yak oocytes and apoptosis[J].Acta Veterinaria et Zootechnica Sinica, 2018, 49(9):1899-1907.(in Chinese)
[49]	字向东, 罗斌, 夏威, 等.基于RNA-Seq技术的牦牛体外受精胚胎发育转录组分析[J].中国农业科学, 2018, 51(8):1577-1589.ZI X D, LUO B, XIA W, et al.Transcriptomic analysis of IVF embryonic development in the yak (Bos grunniens) via RNA-Seq[J].Scientia Agricultura Sinica, 2018, 51(8):1577-1589.(in Chinese)

Cloning and Analysis of Bioinformatics and Tissue Expression Characteristics of Yak StAR Gene

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 49

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LUO Ting, HAN Zhu, XU Yefen, CAI Lin, SUOLANG Sizhu, XU Jinhua, NIU Jiaqiang. Whole Genome Sequencing and Sequence Analysis on T10 of Mycoplasma bovis Strain from Yaks in Xizang [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2154-2167.
[2]	HUANG Xianpeng, XING Jiayi, BAI Yuanyuan, JIANG Yuting, MA Zhiwei, FU Wei, LAN Daoliang. Cloning of Six Pluripotent Related Transcription Factors OSKMNL in Yak and Construction of Polycistron Lentiviral Vector [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1579-1591.
[3]	SHANG Kaiyuan, JIANG Mingfeng, GUAN Jiuqiang, AN Tianwu, ZHAO Hongwen, BAI Qin, WU Weisheng, LI Huade, XIE Rongqing, SHA Quan, LUO Xiaolin, ZHANG Xiangfei. Effects of Maternal Nutritional Regulation in Transition Period on Growth and Development, Serum Biochemistry and Immune Function of Yak Calves [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1638-1648.
[4]	ZUO Zizhen, WANG Haibo, CHAI Zhixin, FU Jianhui, ZHANG Xiangfei, LUO Xiaolin, ZHONG Jincheng. Effects of Rumen-protected Methionine on Meat Quality, Volatile Flavor Compounds and Fatty Acid Composition of Yak Semitendinosus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1102-1114.
[5]	LIU Bin, WANG Meng, PAN Yangyang, WANG Jinglei, XU Gengquan. Effect of LPA on the Expression of HAS2, PTGS2 and PTX3 in Cumulus Cells of Yak (Bos grunniens) [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 552-561.
[6]	CHEN Mengjuan, LIU Yuqing, WANG Zhitong, WEN Jiale, XU Huifen, YU Guangqing, LI Ming. Construction of Eukaryotic Expression Vector, Expression Pattern of BMP15 Gene, and Its Expression in Ovary of New Zealand White Rabbit [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 562-575.
[7]	LIU Yili, TANG Jiao, MIN Qi, YANG Lu, WANG Zening, HU Lian, ZHAO Di, JIANG Mingfeng. Mining Key Candidate Genes of Development and Metabolism in Yak Abomasum Based on Transcriptome Data [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 153-168.
[8]	YAO Ying, ZHOU Yingcong, DU Peiyan, LI Yijuan, QIAN Wenjie, LI Liuyang, YU Zhipeng, CUI Yan, YU Sijiu, FAN Jiangfeng. Proteomic Analysis of Yak Serum During Pregnancy Based on TMT Technology [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 192-206.
[9]	ZHANG Ying, YUAN Ligang, CHEN Guojuan, ZHANG Fang, YANG Dapeng. Analysis of Differential Expression of ET-1/eNOS in the Development of Cryptorchidism in Yak [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 207-217.
[10]	LI Zaiwen, LI Xiang, LI Xiaowei, LI Biao, JIANG Mingfeng. The Pedigree Reconstruction of the Maiwa Yak Preserved Population Based on GBS Technology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3710-3721.
[11]	WANG Jingyu, PAN Yangyang, XU Gengquan, ZHANG Rui, ZHANG Wenlan, WANG Xiaoshan, WU Rentaodi, ZHAO Rigetu, CUI Yan, YU Sijiu. Preparation and Preliminary Application of Yak (Bos Grunniens) Fas-associated Factor 1 Polyclonal Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3369-3382.
[12]	ZHANG Peng, WANG Mingxiu, JING Kemin, PENG Wei, TIAN Yuan, LI Yuqian, FU Changqi, SHU Shi, ZHONG Jincheng, CAI Xin. Abnormal Expression of FGFs/FGFRs and Their Mediated Signaling Pathway Genes Affect the Proliferative Activity of Undifferentiated Spermatogonia in Cattleyak [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2886-2897.
[13]	SHAO Yuexin, ZHANG Xinyu, GE Liyan, SHI Huaiping. Cloning and RNA Interference Analysis of ATF4 Gene in Xinong Saanen Dairy Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2353-2364.
[14]	JIAO Zhengxing, PAN Yangyang, WANG Meng, WANG Jinglei, MA Wenbin, GAO Xiang, ZHANG Hui, CUI Yan, YU Sijiu, WANG Libin. Preparation of Polyclonal Antibody against Yak LC3B Protein and Its Application in Detection of Expression in Reproductive Organs [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2436-2447.
[15]	CHANG Yiming, TANG Cheng, YUE Hua. Molecular Epidemiological Investigation of Bovine Respiratory Syncytial Virus in Yaks [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2030-2041.