表达猫杯状病毒NS2质粒转染猫肾传代细胞的转录组学分析

doi:10.11843/j.issn.0366-6964.2025.12.036

Abstract

Abstract: This study aims to investigate the interaction between FCV NS2 protein and host cells. In this study, a eukaryotic expression plasmid of FCV NS2 was transfected into CRFK cells, with an empty vector as the control. Three samples from each group were collected 24 hours post-transfection for transcriptomic sequencing. Results were as follows: Comparison of the sequencing results revealed 218 differentially expressed genes (DEGs) in the experimental group transfected with NS2 protein, including 200 upregulated genes and 18 downregulated genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the significantly differentially expressed genes were mainly involved in intracellular viral invasion and immune evasion pathways. This study reveals that the NS2 protein significantly regulates multiple signaling pathways at the transcriptional level, including endosomal transport, TGF-β, and RIG-I, in CRFK cells, systematically reveals the global transcriptional regulation patterns of host cells by feline calicivirus (FCV) NS2 protein. These findings provide a new perspective for future research on the interactions between FCV and host cells.

Key words: feline calicivirus, NS2, transcriptome sequencing, differentially expressed genes

CLC Number:

S852.659.6

CHEN Shangchen, DONG Ningning, TAN Xiaomei, LI Na, ZHANG Qi, LUO Maoqing, DENG Yingkun, PANG Shuaisai, GAO Jiaxi, LIU Guangqing, MENG Chunchun. Differential Transcriptomic Analysis of Feline Kidney Cells Transfected with Feline Calicivirus NS2 Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6339-6350.

References

[1] PESAVENTO P A, CHANG K O, PARKER J S. Molecular virology of feline calicivirus [J]. Vet Clin North Am Small Anim Pract, 2008, 38(4): 775-786.
[2] SMITH S L, AFONSO M M, PINCHBECK G L, et al. Temporally separated feline calicivirus isolates do not cluster phylogenetically and are similarly neutralised by high-titre vaccine strain FCV-F9 antisera in vitro [J]. J Feline Med Surg, 2020, 22(6): 602-607.
[3] HERBERT T P, BRIERLEY I, BROWN T D. Identification of a protein linked to the genomic and subgenomic mRNAs of feline calicivirus and its role in translation [J]. J Gen Virol, 1997, 78(Pt 5): 1033-1040.
[4] BAILEY D, KAISER W J, HOLLINSHEAD M, et al. Feline calicivirus p32, p39 and p30 proteins localize to the endoplasmic reticulum to initiate replication complex formation [J]. J Gen Virol, 2010, 91(Pt 3): 739-749.
[5] NATONI A, KASS G E N, CARTER M J, et al. The mitochondrial pathway of apoptosis is triggered during feline calicivirus infection [J]. J Gen Virol, 2006, 87(Pt 2): 357-361.
[6] 蒋婷, 李文东, 李兴起, 等. 转录组和蛋白组筛选就巢鸡卵巢发育候选基因及其调控网络构建 [J]. 畜牧兽医学报, 2024, 55(11): 4950-4967.
JIANG T, LI W D, LI X Q, et al. Screening candidate genes for ovarian development and constructing regulatory network in nesting chickens by transcriptome and proteome [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 4950-4967. (in Chinese)
[7] 朱靓芸, 曾一桐, 童庆芳, 等. 猪流行性腹泻病毒感染猪肾细胞的代谢组学研究 [J]. 中国兽医科学, 2023, 53(8): 1065-1072.
ZHU L Y, ZENG Y T, TONG Q F, et al. Metabonomics of porcine kidney cells infected with porcine epidemic diarrhea virus [J]. Veterinary Science in China, 2023, 53(8): 1065-1072. (in Chinese)
[8] 陈晶晶. Sirt3调控小胶质细胞活化在血管性认知障碍中的作用及机制研究 [D]. 上海：中国人民解放军海军军医大学, 2024.
CHEN J J. The Function and mechanism of Sirt3 in regulating microglial activation during vascular cognitive impairment [D]. Shanghai：Naval Medical University, 2024. (in Chinese)
[9] YUMIKETA Y, NARITA T, INOUE Y, et al. Nonstructural protein p39 of feline calicivirus suppresses host innate immune response by preventing IRF-3 activation [J]. Vet Microbiol, 2016, 185: 62-67.
[10] KAISER W J, CHAUDHRY Y, SOSNOVTSEV S V, et al. Analysis of protein-protein interactions in the feline calicivirus replication complex [J]. J Gen Virol, 2006, 87(Pt 2): 363-8.
[11] CHEN P-I, SCHAUER K, KONG C, et al. Rab5 isoforms orchestrate a "division of labor" in the endocytic network; Rab5C modulates Rac-mediated cell motility [J]. PloS One, 2014, 9(2): e90384.
[12] CHRISTOFORIDIS S, MCBRIDE H M, BURGOYNE R D, et al. The Rab5 effector EEA1 is a core component of endosome docking [J]. Nature, 1999, 397(6720): 621-625.
[13] ZHAO Z, WEI Z, ZHENG J, et al. Hepatitis B virus promotes its own replication by enhancing RAB5A-mediated dual activation of endosomal and autophagic vesicle pathways [J]. Emerg Microbes Infect, 2023, 12(2): 2261556.
[14] ANDRÁS I E, TOBOREK M. HIV-1 stimulates nuclear entry of amyloid beta via dynamin dependent EEA1 and TGF-β/Smad signaling [J]. Exp Cell Res, 2014, 323(1): 66-76.
[15] TOPOL L Z, MARX M, LAUGIER D, et al. Identification of drm, a novel gene whose expression is suppressed in transformed cells and which can inhibit growth of normal but not transformed cells in culture [J]. Mol Cell Biol, 1997, 17(8): 4801-4810.
[16] CHEN B, BLAIR D G, PLISOV S, et al. Cutting edge: bone morphogenetic protein antagonists Drm/Gremlin and Dan interact with slits and act as negative regulators of monocyte chemotaxis [J]. J Immunol, 2004, 173(10): 5914-5917.
[17] FRAMPTON A R, UCHIDA H, VON EINEM J, et al. Equine herpesvirus type 1 (EHV-1) utilizes microtubules, dynein, and ROCK1 to productively infect cells [J]. Vet Microbiol, 2010, 141(1): 12-21.
[18] KUMAR R, CHANDER Y, KHANDELWAL N, et al. ROCK1/MLC2 inhibition induces decay of viral mRNA in BPXV infected cells [J]. Sci Rep, 2022, 12(1): 17811.
[19] HARMEGNIES D, WANG X-M, VANDENBUSSCHE P, et al. Characterization of a potent human interleukin-11 agonist [J]. Biochem J, 2003, 375(Pt 1): 23-32.
[20] LAMERTZ L, RUMMEL F, POLZ R, et al. Soluble gp130 prevents interleukin-6 and interleukin-11 cluster signaling but not intracellular autocrine responses [J]. Sci Signal, 2018, 11(550).
[21] SHARIQ M, MALIK A A, SHEIKH J A, et al. Regulation of autophagy by SARS-CoV-2: The multifunctional contributions of ORF3a [J]. J Med Virol, 2023, 95(7): e28959.
[22] KIM M J, HWANG S Y, IMAIZUMI T, et al. Negative feedback regulation of RIG-I-mediated antiviral signaling by interferon-induced ISG15 conjugation [J]. J Virol, 2007, 82: 1474-1483.
[23] RODRIGUEZ MARISELA R, MONTE K, THACKRAY LARISSA B, et al. ISG15 functions as an interferon-mediated antiviral effector early in the murine norovirus life cycle [J]. J Virol, 2014, 88(16): 9277-9286.
[24] ZHAO C, SRIDHARAN H, CHEN R, et al. Influenza B virus non-structural protein 1 counteracts ISG15 antiviral activity by sequestering ISGylated viral proteins [J]. Nat Commun, 2016, 7(1): 12754.

Differential Transcriptomic Analysis of Feline Kidney Cells Transfected with Feline Calicivirus NS2 Protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIN Xiao, LI Ruijie, LIU Long, GENG Tuoyu, GONG Daoqing. Research Progress on Sex Determining Genes and Their Methylation Regulation in Animals [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4129-4142.
[2]	FAN Jing, LI Wei, ZHU Yan, Wudubala , SHI Jiahui, Husile , WU Jianghong. Study on Rumen Morphological Changes and Gene Expression Differences in Hu Sheep at Different Developmental Stages [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3773-3786.
[3]	LI Bowen, WANG Shujuan, HU Yufeng, WANG Yibing, GAN Xueqiang, CHEN Yanyan, WANG Dongfang, MA Zhenyuan, ZHAO Xueli, YAN Ruoqian. Isolation and Identification of Feline Calicivirus and Analysis of Important Nucleotide Sequences [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 4112-4119.
[4]	SU Meng, LIU Sha, SONG Danli, GAO Qianmei, ZHENG Maiqing, WEN Jie, ZHAO Guiping, LI Qinghe. Identification of Candidate Genes Associated with Ascites Syndrome in Broilers Based on Transcriptome Sequencing [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 559-570.
[5]	QIAO Liying, XU Changsong, ZHANG Li, DING Yi, PAN Yangyang, YANG Kaijie, LIU Jianhua, LIU Wenzhong. Study on Function of CTCF Regulating the Differentiation of Ovine Preadipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6130-6144.
[6]	HE Siqi, CHEN Qian, JIANG Lin, MA Yuehui, ZHOU Shenghua, ZHAO Qianjun. The Effect of METTL14 on Myogenic Differentiation of Ovine Skeletal Muscle Satellite Cells Based on Transcriptome Sequencing Analysis [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 4925-4937.
[7]	Xiaoxu ZHANG, Hao LI, Pingjie FENG, Hao YANG, Xinyue LI, Ran LÜ, Zhangyuan PAN, Mingxing CHU. Application of Single-Cell Transcriptome Sequencing Technology in Domesticated Animals [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3276-3287.
[8]	Mingliang HE, Xiaoyang LÜ, Yongqing JIANG, Zhenghai SONG, Yeqing WANG, Huiguo YANG, Shanhe WANG, Wei SUN. Function Analysis of SOX18 in Hu Sheep Hair Follicle Dermal Papilla Cells Based on Transcriptome Sequencing [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2409-2420.
[9]	WANG Xin, NIE Tong, LI Aqun, MA Jun. Hesperidin Alleviates High-fat-diet Induced Hepatic Oxidative Stress in Mice via Oxidative Phosphorylation Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1302-1313.
[10]	GAO Yawei, PENG Di, SUN Zhaoyang, YAN Ziyue, CUI Kai, MA Zefang. Mining the Molecular Mechanism of Exogenous Melatonin Affecting the Development of Mink Ovary Based on Transcriptome Data [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 607-618.
[11]	MIN Xiangyu, WEI Jiali, XU Biao, LIU Huitao, ZHENG Junjun, WANG Guiwu. Full-length Transcriptome Sequencing of Sika Deer Antler and Mining of Antler Yield-related Genes [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5549-5566.
[12]	ZHANG Zeyu, DONG Ningning, TAN Xiaomei, LI Chuanfeng, ZHU Jie, LIU Guangqing, ZHANG Wei, MENG Chunchun. Preparation of Monoclonal Antibodies against Feline Calicivirus VP1 Protein and Identification of Antigenic Epitopes [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5784-5791.
[13]	Yinliang ZHANG, Ran ZHANG, Wenjun WANG, Dehe WANG, Lanhui LI, Rongyan ZHOU. Mining of Key Candidate Genes Involved in Bone Metabolism Differences at Pre- and Post-laying Stage Based on Transcriptome Data in Laying Hens [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4455-4465.
[14]	WANG Ying, LI Jiakang, ZENG Yue, SHI Kaituo, WEI Liting, PENG Jiajia, LI Qiuyan, CAO Longlong, ZHOU Dengyuan. Development of Monoclonal Antibodies against Feline Calicivirus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 401-405.
[15]	HU Ting, ZHANG Yonghong, HOU Xiaolin, YAO Hua, CUI Defeng, PAN Zaozao, ZHANG Lingyu, ZHANG Jiaxi, WU Qiong. The Effects of Bisphenol A on Inflammation and Amino Acid Metabolism Pathways in Porcine Testis Sertoli Cells Based on Transcriptome Analysis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2858-2871.