H9N2亚型流感病毒感染过程中神经介素B及受体对NF-κB信号通路泛素酶的调控

doi:10.11843/j.issn.0366-6964.2023.07.042

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (7): 3118-3126.doi: 10.11843/j.issn.0366-6964.2023.07.042

• 研究简报 • 上一篇

H9N2亚型流感病毒感染过程中神经介素B及受体对NF-κB信号通路泛素酶的调控

田世茂, 万乾晖, 许晓东, 孔迎迎, 田珂, 唐钰冰, 陈吉龙, 杨桂红*

福建农林大学动物科学学院(蜂学学院) 闽台动物病原生物学重点实验室, 福州 350002

收稿日期:2022-11-30 出版日期:2023-07-23 发布日期:2023-07-21
通讯作者: 杨桂红,主要从事小肽在动物疾病感染过程中的作用研究,E-mail:yangguihong@fafu.edu.cn
作者简介:田世茂(1998-),女,贵州贵阳人,硕士生,主要从事基础兽医学研究,E-mail:tianshimao@fafu.edu.cn
基金资助:
国家自然科学基金重点项目（32030110）；福建省自然科学基金面上项目（2021J01085/2020J01539）；福建农林大学科技专项创新基金项目（CXZX2020060A）

Ubiquitinase of NF-κB Signal Pathway Regulated by Neuromedin B and Its Receptor NMBR during Influenza A Virus H9N2 Subtype Infection

TIAN Shimao, WAN Qianhui, XU Xiaodong, KONG Yingying, TIAN Ke, TANG Yubing, CHEN Jilong, YANG Guihong*

Key Laboratory of Fujian-Taiwan Animal Pathogen Biology, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China

Received:2022-11-30 Online:2023-07-23 Published:2023-07-21

摘要/Abstract

摘要： 神经介素B （neuromedin B，NMB）及受体（NMB receptor，NMBR）通过NF-κB信号通路参与抑制甲型流感病毒（influenza A virus，IAV） H1N1亚型（IAV/H1N1）的感染。但关于NMB和NMBR对NF-κB信号通路相关泛素蛋白酶的调控如何，尚未见报道。为探究NMB和NMBR调控IAV诱导的NF-κB信号通路相关的泛素蛋白酶表达的影响，本研究基于IAV/H9N2感染sh-NMBR细胞与NMB刺激的A549细胞，采用RT-PCR、qRT-PCR及Western blot （WB）分析NMB和NMBR对H9N2感染引起的NF-κB信号通路相关的E3泛素连接酶Mind bomb-2（MIB2）和Ring Finger Protein 8（RNF8）及去泛素蛋白酶Cylindromatosis （CYLD）的表达变化。结果显示：H9N2感染sh-NMBR细胞中，RNF8和CYLD表达增加，MIB2表达和P65磷酸化水平降低；NMB激活A549细胞中NMBR的表达后，诱导细胞中RNF8和CYLD的表达水平下降，MIB2表达和P65磷酸化水平增加。结果表明：NMB和NMBR通过调控IAV/H9N2感染诱导的泛素蛋白酶的表达和P65活性，从而影响IAV/H9N2感染激活的NF-κB信号通路的功能，该结果为深入研究NMB和NMBR抑制甲型流感病毒感染的作用机制提供了理论基础。

关键词: 神经介素B, 神经介素B受体, IAV/H9N2亚型, E3泛素连接酶, P65磷酸化

Abstract: Neuromedin B(NMB) and its receptor(NMBR) could inhibit the infection of influenza A virus(IAV) H1N1 subtype through activating the pathway of NF-κB signaling. However, the regulation of NMB and NMBR on the expression of ubiquitination ligases related with NF-κB signaling pathway remains unclear. To explore the effects of NMB and NMBR on regulating the ubiquitination ligases involved in NF-κB signaling pathway during IAV/H9N2 infection, the expression levels of E3 ubiquitin ligase mind bomb-2(MIB2) and ring finger protein 8(RNF8), deubiquitin enzyme cylindromatosis(CYLD) involved in NF-κB signaling pathway were analyzed using the methods of RT-PCR, qRT-PCR, and Western blot(WB), based on the sh-NMBR cells and NMB stimulating A549 cells during IAV/H9N2 infection. The results showed that the increased expression levels of RNF8 and CYLD, and the decreased levels of MIB2 and P65 phosphorylation were observed in sh-NMBR cells during H9N2 infection, while the decreased levels of RNF8 and CYLD, and the increased levels of MIB2 and P65 phosphorylation were confirmed in NMB stimulating A549 cells. These results indicated that NMB and NMBR could affect the function of NF-κB signaling pathway by regulating the expression levels of ubiquitination ligases and P65 phosphorylation during IAV/H9N2 infection, which could provide a theoretical basis for further studying the mechanism of NMB and NMBR on inhibiting influenza A virus.

Key words: neuromedin B, neuromedin B receptor, influenza A virus H9N2 subtype, E3 ubiquitination enzymes, P65 phosphorylation

中图分类号:

R511.7

田世茂, 万乾晖, 许晓东, 孔迎迎, 田珂, 唐钰冰, 陈吉龙, 杨桂红. H9N2亚型流感病毒感染过程中神经介素B及受体对NF-κB信号通路泛素酶的调控[J]. 畜牧兽医学报, 2023, 54(7): 3118-3126.

TIAN Shimao, WAN Qianhui, XU Xiaodong, KONG Yingying, TIAN Ke, TANG Yubing, CHEN Jilong, YANG Guihong. Ubiquitinase of NF-κB Signal Pathway Regulated by Neuromedin B and Its Receptor NMBR during Influenza A Virus H9N2 Subtype Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3118-3126.

参考文献 31

[1]	TE VELTHUIS A J W, FODOR E. Influenza virus RNA polymerase:insights into the mechanisms of viral RNA synthesis[J]. Nat Rev Microbiol, 2016, 14(8):479-493.
[2]	MEDINA R A, GARCÍA-SASTRE A. Influenza A viruses:new research developments[J]. Nat Rev Microbiol, 2011, 9(8):590-603.
[3]	JIMENEZ-BLUHM P, SEPULVEDA A, BAUMBERGER C, et al. Evidence of influenza infection in dogs and cats in central Chile[J]. Prev Vet Med, 2021, 191:105349.
[4]	DAI M L, DU W J, MARTÍNEZ-ROMERO C, et al. Analysis of the evolution of pandemic influenza A(H1N1) virus neuraminidase reveals entanglement of different phenotypic characteristics[J]. mBio, 2021, 12(3):e00287-21.
[5]	CHEN R B, HOLMES E C. Avian influenza virus exhibits rapid evolutionary dynamics[J]. Mol Biol Evol, 2006, 23(12):2336-2341.
[6]	WU J, GU J Q, SHEN L, et al. The role of host cell Rab GTPases in influenza A virus infections[J]. Future Microbiol, 2021, 16:445-452.
[7]	WANG Y C, ZHANG X J, BI K F, et al. Critical role of microRNAs in host and influenza A (H1N1) virus interactions[J]. Life Sci, 2021, 277:119484.
[8]	NENASHEVA V V, NIKITENKO N A, STEPANENKO E A, et al. Human TRIM14 protects transgenic mice from influenza A viral infection without activation of other innate immunity pathways[J]. Genes Immun, 2021, 22(1):56-63.
[9]	LANZ C, SCHOTSAERT M, MAGNUS C, et al. IFITM3 incorporation sensitizes influenza A virus to antibody-mediated neutralization[J]. J Exp Med, 2021, 218(6):e20200303.
[10]	SUN Y P, LIU J H. H9N2 influenza virus in China:a cause of concern[J]. Protein Cell, 2015, 6(1):18-25.
[11]	BI Y H, CHEN Q J, WANG Q L, et al. Genesis, evolution and prevalence of H5N6 avian influenza viruses in China[J]. Cell Host Microbe, 2016, 20(6):810-821.
[12]	KAMEDA H, MIYOSHI H, SHIMIZU C, et al. Expression and regulation of neuromedin B in pituitary corticotrophs of male melanocortin 2 receptor-deficient mice[J]. Endocrinology, 2014, 155(7):2492-2499.
[13]	YANG G H, HUANG H P, TANG M Y, et al. Role of neuromedin B and its receptor in the innate immune responses against influenza A virus infection in vitro and in vivo[J]. Vet Res, 2019, 50(1):80.
[14]	唐梦瑶, 马逸杰, 田世茂, 等. 神经介素B及其受体NMBR参与抗A型流感病毒H1N1亚型感染的信号通路[J]. 畜牧兽医学报, 2021, 52(11):3215-3223.TANG M Y, MA Y J, TIAN S M, et al. The signaling pathway of neuromedin B and its receptor NMBR involvement in anti-influenza A virus H1N1 subtype infection[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11):3215-3223. (in Chinese)
[15]	DAI M M, LI S B, SHI K Y, et al. Comparative analysis of key immune protection factors in H9N2 avian influenza viruses infected and immunized specific pathogen-free chicken[J]. Poult Sci, 2021, 100(1):39-46.
[16]	LIU S, ZHUANG Q Y, WANG S C, et al. Control of avian influenza in China:strategies and lessons[J]. Transbound Emerg Dis, 2020, 67(4):1463-1471.
[17]	HAYDEN M S, GHOSH S. NF-κB in immunobiology[J]. Cell Res, 2011, 21(2):223-244.
[18]	SCHEIDEREIT C. IκB kinase complexes:gateways to NF-κB activation and transcription[J]. Oncogene, 2006, 25(51):6685-6705.
[19]	KRAPPMANN D, SCHEIDEREIT C. A pervasive role of ubiquitin conjugation in activation and termination of IκB kinase pathways[J]. EMBO Rep, 2005, 6(4):321-326.
[20]	HINZ M, SCHEIDEREIT C. The IκB kinase complex in NF-κB regulation and beyond[J]. EMBO Rep, 2014, 15(1):46-61.
[21]	AFONINA I S, ZHONG Z Y, KARIN M, et al. Limiting inflammation-the negative regulation of NF-κB and the NLRP3 inflammasome[J]. Nat Immunol, 2017, 18(8):861-869.
[22]	ZHANG Q, LENARDO M J, BALTIMORE D. 30 Years of NF-κB:a blossoming of relevance to human pathobiology[J]. Cell, 2017, 168(1-2):37-57.
[23]	SHI H X, SUN L, WANG Y, et al. N4BP1 negatively regulates NF-κB by binding and inhibiting NEMO oligomerization[J]. Nat Commun, 2021, 12(1):1379.
[24]	GAO S J, WU J X, LIANG L L, et al. RNF8 negatively regulates NF-kappaB signaling by targeting IkappaB kinase:implications for the regulation of inflammation signaling[J]. Biochem Biophys Res Commun, 2017, 488(1):189-195.
[25]	UEMATSU A, KIDO K, TAKAHASHI H, et al. The E3 ubiquitin ligase MIB2 enhances inflammation by degrading the deubiquitinating enzyme CYLD[J]. J Biol Chem, 2019, 294(38):14135-14148.
[26]	SUN S C. The non-canonical NF-κB pathway in immunity and inflammation[J]. Nat Rev Immunol, 2017, 17(9):545-558.
[27]	HAYDEN M S, GHOSH S. Shared principles in NF-κB signaling[J]. Cell, 2008, 132(3):344-362.
[28]	HU H B, SUN S C. Ubiquitin signaling in immune responses[J]. Cell Res, 2016, 26(4):457-483.
[29]	CHEN Z J, PARENT L, MANIATIS T. Site-specific phosphorylation of IκBα by a novel ubiquitination-dependent protein kinase activity[J]. Cell, 1996, 84(6):853-862.
[30]	YAMAOKA S, COURTOIS G, BESSIA C, et al. Complementation cloning of NEMO, a component of the IκB kinase complex essential for NF-κB activation[J]. Cell, 1998, 93(7):1231-1240.
[31]	ISRAËL A. The IKK complex, a central regulator of NF-κB activation[J]. Cold Spring Harb Perspect Biol, 2010, 2(3):a000158.

H9N2亚型流感病毒感染过程中神经介素B及受体对NF-κB信号通路泛素酶的调控

Ubiquitinase of NF-κB Signal Pathway Regulated by Neuromedin B and Its Receptor NMBR during Influenza A Virus H9N2 Subtype Infection

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[1]	唐梦瑶, 马逸杰, 田世茂, 万乾晖, 杨桂红. 神经介素B及其受体NMBR参与抗A型流感病毒H1N1亚型感染的信号通路[J]. 畜牧兽医学报, 2021, 52(11): 3215-3223.
[2]	陈轶霞，邵忠伟，李贵华，王聪. 具有E3泛素连接酶功能的痘病毒蛋白质[J]. 畜牧兽医学报, 2015, 46(12): 2127-2134.