鸡PERP1基因功能分析、核心启动子筛选及其转录因子预测

doi:10.16431/j.cnki.1671-7236.2023.09.001

Abstract

Abstract: 【Objective】 The purpose of this study was to investigate the regulatory role and expression mechanism of TP53 apoptotic effector factor (PERP1) in follicular development.【Method】 Using ovarian tissue of laying hens as material,the full length sequence of the CDS region of PERP1 gene in laying hens was cloned,and an overexpression vector of the PERP1 gene was constructed.The relative expression of PERP1 gene,proliferation and apoptosis related genes BCL2 and c-Myc,and oxidative stress related genes SOD2 and CAT after transfection were detected using Real-time quantitative PCR.Three online bioinformatics analysis software were used to predict the core promoter region of PERP1 gene in chicken,and six promoter deletion primers were designed according to the prediction results.Using blood tissues of laying hens as materials,six promoter deletion fragments of PERP1 gene 5 '-UTR were cloned and recombinant plasmids were constructed,and the location of the promoter active region was verified by Luciferase reporter gene test.Use four transcription factor online prediction software to predict and analyze transcription factors in the promoter active region.【Result】 This study successfully obtained the full length sequence of the CDS region of PERP1 gene in chicken and constructed an overexpression vector for PERP1 gene.After overexpression of PERP1 gene, compared with control group, the anti apoptotic gene BCL2 and proliferative gene c-Myc were significantly downregulated (P＜0.01),while the relative expression of antioxidant stress genes CAT and SOD2 were not significantly different (P＞0.05).The results of bioinformatics software prediction and Luciferase report vector test showed that the core promoter of PERP1 gene was located upstream of the CDS region (―441/―71 bp).The transcription factor prediction results revealed 13 candidate transcription factors,including Sp1,ZEB1,Zic3,c-Jun,AP-2alpha,AP-1,NF-1,c/EBPalp,Adf-1,HNF-3,CACCC-bi,c-Myc,and Smad4.【Conclusion】 The PERP1 gene had the function of promoting granulosa cell apoptosis,and regulating its expression was of great significance for follicular development.

Key words: chickens; PERP1 gene; core promoter; granulosa cell; transcription factors

CLC Number:

S831.2
Q78

CHEN Lin, WANG Jiaxiang, WU Yan, PI Jinsong, ZHANG Ying, LI Chengfeng. Functional Analysis, Core Promoter Screening and Transcription Factors Prediction of PERP1 Gene in Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3449-3458.

References

[1] JOHNSON P A.Follicle selection in the avian ovary [J]. Reproduction in Domestic Animals,2012,47(Suppl 4):283-287.
[2] JIN Y,ZHANG C,LIN X,et al.Prostaglandin involvement in follicle-stimulating hormone-induced proliferation of granulosa cells from chicken prehierarchical follicles [J].Prostaglandins & Other Lipid Mediators,2006,81(1-2):45-54.
[3] MCELROY A P,CALDWELL D J,PROUDMAN J A,et al.Modulation of in vitro DNA synthesis in the chicken ovarian granulosa cell follicular hierarchy by follicle-stimulating hormone and luteinizing hormone [J].Poultry Science,2004,83(3):500-506.
[4] ZHOU S,MA Y,YAO J,et al.TGF-β1-induced collagen promotes chicken ovarian follicle development via an intercellular cooperative pattern [J]. Cell Biology International,2021,45(6):1336-1348.
[5] LIN X,MA Y,QIAN T,et al.Basic fibroblast growth factor promotes prehierarchical follicle growth and yolk deposition in the chicken [J]. Theriogenology,2019,139:90-97.
[6] LIN J X,JIA Y D,ZHANG C Q.Effect of epidermal growth factor on follicle-stimulating hormone-induced proliferation of granulosa cells from chicken prehierarchical follicles [J].Journal of Zhejiang University-Science B,2011,12(11):875-883.
[7] SUN X,NIU X,QIN N,et al.Novel insights into the regulation of LATS2 kinase in prehierarchical follicle development via the Hippo pathway in hen ovary [J].Poultry Science,2021,100(12):101454.
[8] LI J,ZHAO D,GUO C,et al.Involvement of Notch signaling in early chick ovarian follicle development [J].Cell Biology International,2016,40(1):65-73.
[9] BEG M A,BERGFELT D R,KOT K,et al.Follicle selection in cattle:Dynamics of follicular fluid factors during development of follicle dominance [J]. Biology of Reproduction,2002,66(1):120-126.
[10] ROBERTS O,PARAOAN L.PERP-ing into diverse mechanisms of cancer pathogenesis:Regulation and role of the p53/p63 effector PERP [J]. Biochimica et Biophysica Acta-Reviews on Cancer,2020,1874(1):188393.
[11] BEAUDRY V G,JIANG D,DUSEK R L,et al.Loss of the p53/p63 regulated desmosomal protein Perp promotes tumorigenesis [J]. PLoS Genetics,2010,6(10):e1001168.
[12] CHEN X,SUN X,CHIMBAKA I M,et al.Transcriptome analysis of ovarian follicles reveals potential pivotal genes associated with increased and decreased rates of chicken egg production [J]. Frontiers in Genetics,2021,12:622751.
[13] LEFAUCHEUR L,ECOLAN P,PLANTARD L,et al.New insights into muscle fiber types in the pig [J]. Journal of Histochemistry & Cytochemistry,2002,50(5):719-730.
[14] JAISHANKAR J,BHATOA L,PATIL N,et al.Microarray profiling and identification of core promoter sequence in Gordonia [J].Genomics,2021,113(6):4327-4336.
[15] LIU R,LIU L,LI X,et al.Engineering yeast artificial core promoter with designated base motifs [J].Microbial Cell Factories,2020,19(1):38.
[16] 林艳丽,覃建兵,伍翔,等.虎杖PcMYB1启动子的克隆及其活性分析[J].生物技术通报,2021,37(5):48-55. LIN Y L,QIN J B,WU X,et al.Cloning and activity analysis of PcMYB1 promoter from Polygonum cuspidatum [J] Biotechnology Bulletin,2021,37(5):48-55.(in Chinese)
[17] 谭光辉,罗华伦,李杰章,等.鸭SCD-1基因启动子的克隆与转录活性分析[J] 基因组学与应用生物学,2020,39(11):5052-5058. TAN G H,LUO H L,LI J Z,et al.Cloning and transcriptional activity analysis of duck SCD-1 gene promoter [J] Genomics and Applied Biology,2020,39(11):5052-5058.(in Chinese)
[18] WANG Y C,LI Z J,HAN R L,et al.Promoter analysis and tissue expression of the chicken ASB15 gene [J].British Poultry Science,2017,58(1):26-31.
[19] YU S,WANG G,LIAO J,et al.A functional mutation in the AMPD1 promoter region affects promoter activity and breast meat freshness in chicken [J]. Animal Genetics,2021,52(1):121-125.
[20] ZHANG Y,HUANG J,LI X,et al.Identification of functional transcriptional binding sites within chicken ABCG2 gene promoter and screening its regulators [J].Genes (Basel),2020,11(2):186.
[21] LI X,LU Y,LIU X,et al.Identification of chicken FSHR gene promoter and the correlations between polymorphisms and egg production in Chinese native hens [J].Reproduction in Domestic Animals,2019,54(4):702-711.
[22] ATTARDI L D,RECZEK E E,COSMAS C,et al.PERP,an apoptosis-associated target of p53,is a novel member of the PMP-22/gas3 family [J]. Genes & Development,2000,14(6):704-718.
[23] DAVIES L,GRAY D,SPILLER D,et al.P53 apoptosis mediator PERP:Localization,function and caspase activation in uveal melanoma [J].Journal of Cellular and Molecular Medicine,2009,13(8b):1995-2007.
[24] KHAN I A,YOO B H,MASSON O,et al.ErbB2-dependent downregulation of a pro-apoptotic protein Perp is required for oncogenic transformation of breast epithelial cells [J].Oncogene,2016,35(44):5759-5769.
[25] 安志芳,魏琳娜,王志洁,等.高原鼢鼠肺组织细胞凋亡相关基因的进化分析及其在不同海拔条件下的表达模式[J].动物学杂志,2019,54(4):549-566. AN Z F,WEI L N,WANG Z J,et al.Evolutionary analysis of apoptosis-related genes in the lung tissue of plateau zokor and their expression patterns at different altitudes [J].Journal of Zoology,2019,54(4):549-566.(in Chinese)
[26] GUO C,ZHANG G,LIN X,et al.Reciprocal stimulating effects of bFGF and FSH on chicken primordial follicle activation through AKT and ERK pathway [J].Theriogenology,2019,132:27-35.
[27] LI J,LUO W,HUANG T,et al.Growth differentiation factor 9 promotes follicle-stimulating hormone-induced progesterone production in chicken follicular granulosa cells[J].General and Comparative Endocrinology,2019,276:69-76.
[28] LAMERS F,SCHILD L,DEN HARTOG I J,et al.Targeted BCL2 inhibition effectively inhibits neuroblastoma tumour growth [J]. European Journal of Cancer,2012,48(16):3093-3103.
[29] HOCKENBERY D M,ZUTTER M,HICKEY W,et al.BCL2 protein is topographically restricted in tissues characterized by apoptotic cell death [J]. Proceedings of the National Academy of Sciences of the United States of America,1991,88(16):6961-6965.
[30] SCHMIDT E V.The role of c-myc in cellular growth control [J].Oncogene,1999,18(19):2988-2996.
[31] SCHMIDT E V.The role of c-myc in regulation of translation initiation [J].Oncogene,2004,23(18):3217-3221.

Functional Analysis, Core Promoter Screening and Transcription Factors Prediction of PERP1 Gene in Chickens

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