小鼠生长激素受体基因环状转录本的克隆及其表达规律

doi:10.16288/j.yczz.21-156

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Hereditas(Beijing) ›› 2021, Vol. 43 ›› Issue (9): 890-900.doi: 10.16288/j.yczz.21-156

• Research Article • Previous Articles Next Articles

Cloning and expression of circular transcript of mouse growth hormone receptor gene

Weilu Zhang(), Qiying Leng, Jiahui Zheng, Ali Hassan Nawaz, Zhenhai Jiao, Fujian Wang, Li Zhang()

College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang 524088, China

Received:2021-04-27 Revised:2021-07-21 Online:2021-09-20 Published:2021-08-19
Contact: Zhang Li E-mail:178052526@qq.com;zhangli761101@163.com
Supported by:
Supported by the National Natural Science Foundation of China Nos(31672412);Supported by the National Natural Science Foundation of China Nos(31972550);the Natural Science Foundation of Guangdong Province No(2020A1515011576);Guangdong-Guangxi Joint Fund No(2020B1515420008)

Abstract

Abstract:

Based on reports in the literature and search results on the circBase database, 8 circular transcripts of the mouse growth hormone receptor (GHR) gene were identified. In order to confirm the existence of the circular transcripts of the GHR gene (circGHRs) and to explore their expression patterns, the Kunming mouse (Mus musculus) was used as a research animal. This study detected the existence of circGHRs by RT-PCR amplification and sequencing, one of which was selected as circGHR for detailed analysis. The circular structure of circGHR was confirmed by RNase R treatment and reverse transcription. The spatiotemporal expression of circGHR and GHR mRNA was analyzed by qRT-PCR. The results showed that the full length of mouse circGHR was 820 nt, which was formed by circularization of exons 2-8 of the transcript of the GHR gene. RNase R tolerance analysis shows that mouse circGHR has the general characteristics of circular molecules and is not easily degraded by RNase R. Compared with oligo-d(T)18 primers, random primers have higher reverse transcription efficiency for circGHR, which further shows that circGHR is a poly(A)-free cyclic structure molecule. Tissue expression profile results show that circGHR is highly expressed in the liver and kidney of 1 week-old and 7-week old Kunming mice, but is low in pectoral muscles and leg muscles. The time-series expression profile of circGHR does not show any significant difference between the liver and pectoral muscle tissue. The circGHR expression in the leg muscle was low before 5 weeks of age but increased after 7 weeks of age. This study confirmed the existence of a circular transcript circGHR of the mouse GHR gene, and initially revealed the expression pattern of circGHR. The results of the study laid a foundation for in-depth developmental studies on the biological functions of the mouse circGHR and its mechanism of action regarding the growth and development of mice.

Key words: mouse, growth hormone receptor, circular RNA

Weilu Zhang, Qiying Leng, Jiahui Zheng, Ali Hassan Nawaz, Zhenhai Jiao, Fujian Wang, Li Zhang. Cloning and expression of circular transcript of mouse growth hormone receptor gene[J]. Hereditas(Beijing), 2021, 43(9): 890-900.

Figures/Tables 9

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References 26

[1]	Palmiter RD, Norstedt G, Gelinas RE, Hammer RE, Brinster RL. Metallothionein-human GH fusion genes stimulate growth of mice. Science, 1983, 222(4625):809-814. pmid: 6356363
[2]	Chen DB, Li QH, Li LH, Tao JG, Chai Y, Wen SP, Wang JH, Liu L, Jia JJ. Growth hormone and growth hormone receptor gene expression. Chin Anim Husb Vet Med, 2007, 34(11):52-54.
	陈宝定, 李琦华, 李丽红, 陶金光, 柴艳, 文生萍, 王金浩, 刘丽, 贾俊静. 生长激素与生长激素受体基因的表达. 中国畜牧兽医, 2007, 34(11):52-54.
[3]	Ma X, Hu Y, Xiong G, He XR, Wang XQ. Overview of growth hormone receptor gene and growth hormone receptor. J Hunan Agric Univ (Nat Sci), 2010, 36(S1):58-62.
	马晓, 胡毅, 熊钢, 何湘蓉, 王晓清. 生长激素受体及其基因研究进展. 湖南农业大学学报(自然科学版), 2010, 36(S1):58-62.
[4]	Argetsinger LS, Carter-Su C. Mechanism of signaling by growth hormone receptor. Physiol Rev, 1996, 76(4):1089-1107. pmid: 8874495
[5]	Luo J, Wang XL, Sun ZC, Wu D, Zhang W, Wang ZJ. Progress in circular RNAs of plants. Hereditas(Beijing), 2018, 40(6):467-477.
	骆甲, 王型力, 孙志超, 吴迪, 张玮, 王正加. 植物环状RNA研究进展. 遗传, 2018, 40(6):467-477.
[6]	Sanger HL, Klotz G, Riesner D, Gross HJ, Kleinschmidt AK. Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures. Proc Natl Acad Sci USA, 1976, 73(11):3852-3856. doi: 10.1073/pnas.73.11.3852
[7]	Jeck WR, Sorrentino JA, Wang K, Slevin MK, Burd CE, Liu JZ, Marzluff WF, Sharpless NE. Circular RNAs are abundant, conserved, and associated with ALU repeats. RNA, 2013, 19(2):141-157. doi: 10.1261/rna.035667.112
[8]	Suzuki H, Zuo YH, Wang JH, Zhang MQ, Malhotra A, Mayeda A. Characterization of RNase R-digested cellular RNA source that consists of lariat and circular RNAs from pre-mRNA splicing. Nucleic Acids Res, 2006, 34(8):e63. doi: 10.1093/nar/gkl151
[9]	Suzuki H, Tsukahara T. A view of pre-mRNA splicing from RNase R resistant RNAs. Int J Mol Sci, 2014, 15(6):9331-9342. doi: 10.3390/ijms15069331
[10]	Chen L, Shan G. Circular RNAs remain peculiarly unclear in biogenesis and function. Sci China Life Sci, 2015, 58(6):616-618. doi: 10.1007/s11427-015-4855-y
[11]	Chen L, Huang C, Wang XL, Shan G. Circular RNAs in eukaryotic cells. Curr Genomics, 2015, 16(5):312-318. doi: 10.2174/1389202916666150707161554
[12]	Huang C, Shan G. What happens at or after transcription: Insights into circRNA biogenesis and function. Transcription, 2015, 6(4):61-64. doi: 10.1080/21541264.2015.1071301
[13]	Liu XQ, Gao YB, Zhao LZ, Cai YC, Wang HY, Miao M, Zhang HX. Biogenesis, research methods, and functions of circular RNAs. Hereditas(Beijing), 2019, 41(6):469-485.
	刘旭庆, 高宇帮, 赵良真, 蔡宇晨, 王汇源, 苗苗, 顾连峰, 张航晓. 环状RNA的产生、研究方法及功能. 遗传, 2019, 41(6):469-485.
[14]	Li ZY, Huang C, Bao C, Chen L, Lin M, Wang XL, Zhong GL, Yu B, Hu WC, Dai LM, Zhu PF, Chang ZX, Wu QF, Zhao Y, Jia Y, Xu P, Liu HJ, Shan G. Exon-intron circular RNAs regulate transcription in the nucleus. Nat Struct Mol Biol, 2015, 22(3):256-264. doi: 10.1038/nsmb.2959
[15]	Legnini I, Di Timoteo G, Rossi F, Morlando M, Briganti F, Sthandier O, Fatica A, Santini T, Andronache A, Wade M, Laneve P, Rajewsky N, Bozzoni I. Circ-ZNF609 is a circular RNA that can be translated and functions in myogenesis. Mol Cell, 2017, 66(1): 22-37.e9. doi: S1097-2765(17)30132-6 pmid: 28344082
[16]	Zheng X, Chen LJ, Zhou Y, Wang Q, Zheng ZJ, Xu B, Wu C, Zhou Q, Hu WW, Wu CP, Jiang JT. A novel protein encoded by a circular RNA circPPP1R12A promotes tumor pathogenesis and metastasis of colon cancer via Hippo-YAP signaling. Mol Cancer, 2019, 18(1):47. doi: 10.1186/s12943-019-1010-6 pmid: 30925892
[17]	Dong R, Ma XK, Chen LL, Yang L. Increased complexity of circRNA expression during species evolution. RNA Biol, 2017, 14(8):1064-1074. doi: 10.1080/15476286.2016.1269999 pmid: 27982734
[18]	Zhen SL, Li L, Zhang HP. Progress on translation ability of circular RNA. Hereditas(Beijing), 2020, 42(5):423-434.
	郑帅龙, 李利, 张红平. 环状RNA翻译能力研究进展. 遗传, 2020, 42(5):423-434.
[19]	Vincent HA, Deutscher MP. Substrate recognition and catalysis by the exoribonuclease RNase R. J Biol Chem, 2006, 281(40):29769-29775. pmid: 16893880
[20]	Rybak-Wolf A, Stottmeister C, Glažar P, Jens M, Pino N, Giusti S, Hanan M, Behm M, Bartok O, Ashwal-Fluss R, Herzog M, Schreyer L, Papavasileiou P, Ivanov A, Öhman M, Refojo D, Kadener S, Rajewsky N. Circular RNAs in the mammalian brain are highly abundant, conserved, and dynamically expressed. Mol Cell, 2015, 58(5):870-885. doi: 10.1016/j.molcel.2015.03.027 pmid: 25921068
[21]	Beyea JA, Sawicki G, Olson DM, List E, Kopchick JJ, Harvey S. Growth hormone (GH) receptor knockout mice reveal actions of GH in lung development. Proteomics, 2006, 6(1):341-348. doi: 10.1002/(ISSN)1615-9861
[22]	Beyea JA, Olson DM, Vandergriend RA, Harvey S. Expression of growth hormone and its receptor in the lungs of embryonic chicks. Cell Tissue Res, 2005, 322(3):379-392. doi: 10.1007/s00441-005-0040-0
[23]	Landau D, Domene H, Flyvbjerg A, Grønbaek H, Roberts CT, Argov S, LeRoith D. Differential expression of renal growth hormone receptor and its binding protein in experimental diabetes mellitus. Growth Horm IGF Res, 1998, 8(1):39-45. pmid: 10990443
[24]	Giani JF, Miquet JG, Muñoz MC, Burghi V, Toblli JE, Masternak MM, Kopchick JJ, Bartke A, Turyn D, Dominici FP. Upregulation of the angiotensin-converting enzyme 2/angiotensin-(1-7)/Mas receptor axis in the heart and the kidney of growth hormone receptor knock-out mice. Growth Horm IGF Res, 2012, 22(6):224-233. doi: 10.1016/j.ghir.2012.08.003
[25]	Wei XF, Wang DM, Liu S, Zhou P. Signal peptide and its application in protein expression. Biotechnol Bull, 2006, (6):38-42.
	韦雪芳, 王冬梅, 刘思, 周鹏. 信号肽及其在蛋白质表达中的应用. 生物技术通报, 2006, (6):38-42.
[26]	Zhou B, Yang H, Yang C, Bao YL, Yang SM, Liu J, Xiao YF. Translation of noncoding RNAs and cancer. Cancer Lett, 2021, 497:89-99. doi: 10.1016/j.canlet.2020.10.002 pmid: 33038492

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引物名称	引物序列(5ʹ→3ʹ)	复性温度(℃)	扩增产物长度(bp)	位置	用途
circGHR1	F：TAGTTTGACCGGGATTCGTGG	59	171	Exon7	circGHR1 环状验证
circGHR1	R: GGAACGACACTTGGTGAATCG	59	171	Exon4	circGHR1 环状验证
circGHR2	F：GGTGAGATCCAGACAACGGA	59	134	Exon8	circGHR2 环状验证
circGHR2	R: GACACTTGGTGAATCGAGGC	59	134	Exon4	circGHR2 环状验证
circGHR3	F：GGGATTCGTGGAGACATCCAA	59	343	Exon7	circGHR3 环状验证
circGHR3	R: GACTGCCAGTGCCAAGGTTA	59	343	Exon2	circGHR3 环状验证
circGHR4	F：GACCGGGATTCGTGGAGACATC	61	422	Exon7	circGHR4 环状验证
circGHR4	R: ACGACACTTGGTGAATCGAGG	61	422	Exon4	circGHR4 环状验证
circGHR5	F：CCATCCCATATGGTGGATCTGT	59	142	Intron1	circGHR5 环状验证
circGHR5	R: ATGGGAAAGGAGGTGATGGC	59	142	Intron1	circGHR5 环状验证
circGHR6	F：GCTGGACCAAAAATGTTTCACTGTT	60	176	Exon6	circGHR6 环状验证
circGHR6	R: CGTTGGCTTTCCCTTTTAGCA	60	176	Exon4,5	circGHR6 环状验证
circGHR7	F：CAGCGAAGTCCTCCGTGTAATA	59	100	Exon8	circGHR7 环状验证
circGHR7	R: CAGGGCATTCTTTCCATTCCTG	59	100	Exon6	circGHR7 环状验证
circGHR8	F: CAGTCACCAGCAGCACATTTT	58	70	Exon2	circGHR8 环状验证
circGHR8	R: AGGTTAAGAAGACCTGACAAAGAT	58	70	Exon2	circGHR8 环状验证

引物名称	引物序列(5'→3')	复性温度(℃)	扩增产物长度(bp)	位置	用途
circGHR-D	F：GGGATTCGTGGAGACATCCAA	59	343	Exon7	环状验证
circGHR-D	R: GACTGCCAGTGCCAAGGTTA	59	343	Exon2	环状验证
circGHR-full	F：GTCTCAGGTATGGATCTTTGTCAGG	57	820	Exon2	全长克隆
circGHR-full	R: CTTCTTCACATGCTTCCAATATGTTC	57	820	Exon8	全长克隆
circGHR-DL	F：GGGATTCGTGGAGACATCCAA	59	343	Exon7	circGHR定量
circGHR-DL	R: GACTGCCAGTGCCAAGGTTA	59	343	Exon2	circGHR定量
GHR-DL	F：GGGTGAGATCCAGACAACGG	57	285	Exon8	GHR mRNA定量
GHR-DL	R: TCACCTCCTCCAACTTCCCT	57	285	Exon11	GHR mRNA定量
GAPDH	F：AGGTTGTCTCCTGCGACTTCA	57	184	Exon6	内参基因定量
GAPDH	R: TGGTCCAGGGTTTCTTACTCC	57	184	Exon7	内参基因定量

Cloning and expression of circular transcript of mouse growth hormone receptor gene

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