MERS-CoV-nsp5 expression in human epithelial BEAS 2b cells attenuates type I interferon production by inhibiting IRF3 nuclear translocation.
Humans
Interferon Regulatory Factor-3
/ metabolism
Middle East Respiratory Syndrome Coronavirus
/ immunology
Epithelial Cells
/ metabolism
Viral Nonstructural Proteins
/ metabolism
Interferon Type I
/ metabolism
Cell Line
Cell Nucleus
/ metabolism
Interferon-beta
/ metabolism
Signal Transduction
/ drug effects
Poly I-C
/ pharmacology
Promoter Regions, Genetic
/ genetics
alpha Karyopherins
/ metabolism
Active Transport, Cell Nucleus
DEAD Box Protein 58
/ metabolism
IRF3
Interferon
KPNA4
MERS-CoV
Nuclear translocation
Journal
Cellular and molecular life sciences : CMLS
ISSN: 1420-9071
Titre abrégé: Cell Mol Life Sci
Pays: Switzerland
ID NLM: 9705402
Informations de publication
Date de publication:
12 Oct 2024
12 Oct 2024
Historique:
received:
04
04
2024
accepted:
17
09
2024
revised:
12
09
2024
medline:
12
10
2024
pubmed:
12
10
2024
entrez:
12
10
2024
Statut:
epublish
Résumé
Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is an enveloped, positive-sense RNA virus that emerged in 2012, causing sporadic cases and localized outbreaks of severe respiratory illness with high fatality rates. A characteristic feature of the immune response to MERS-CoV infection is low type I IFN induction, despite its importance in viral clearance. The non-structural proteins (nsps) of other coronaviruses have been shown to block IFN production. However, the role of nsp5 from MERS-CoV in IFN induction of human respiratory cells is unclear. In this study, we elucidated the role of MERS-CoV-nsp5, the viral main protease, in modulating the host's antiviral responses in human bronchial epithelial BEAS 2b cells. We found that overexpression of MERS-CoV-nsp5 had a dose-dependent inhibitory effect on IFN-β promoter activation and cytokine production induced by HMW-poly(I:C). It also suppressed IFN-β promoter activation triggered by overexpression of key components in the RIG-I-like receptor (RLR) pathway, including RIG-I, MAVS, IKK-ε and IRF3. Moreover, the overexpression of MERS-CoV-nsp5 did not impair expression or phosphorylation of IRF3, but suppressed the nuclear translocation of IRF3. Further investigation revealed that MERS-CoV-nsp5 specifically interacted with IRF3. Using docking and molecular dynamic (MD) simulations, we also found that amino acids on MERS-CoV-nsp5, IRF3, and KPNA4 may participate in protein-protein interactions. Additionally, we uncovered protein conformations that mask the nuclear localization signal (NLS) regions of IRF3 and KPNA4 when interacting with MERS-CoV-nsp5, suggesting a mechanism by which this viral protein blocks IRF3 nuclear translocation. Of note, the IFN-β expression was restored after administration of protease inhibitors targeting nsp5, indicating this suppression of IFN-β production was dependent on the enzyme activity of nsp5. Collectively, our findings elucidate a mechanism by which MERS-CoV-nsp5 disrupts the host's innate antiviral immunity and thus provides insights into viral pathogenesis.
Identifiants
pubmed: 39395053
doi: 10.1007/s00018-024-05458-y
pii: 10.1007/s00018-024-05458-y
doi:
Substances chimiques
Interferon Regulatory Factor-3
0
IRF3 protein, human
0
Viral Nonstructural Proteins
0
Interferon Type I
0
Interferon-beta
77238-31-4
KPNA4 protein, human
0
Poly I-C
O84C90HH2L
alpha Karyopherins
0
DEAD Box Protein 58
EC 3.6.4.13
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
433Subventions
Organisme : China Scholarship Council
ID : 201908300032
Organisme : Science Foundation Ireland
ID : SFI 20/SPP/3685
Pays : Ireland
Organisme : Science Foundation Ireland
ID : SFI 19/FFP/6483
Pays : Ireland
Organisme : Irish Research Council
ID : GOIPG/2021/954
Informations de copyright
© 2024. The Author(s).
Références
Zhang Y, Gargan S, Lu Y, Stevenson NJ (2021) An overview of current knowledge of Deadly CoVs and their interface with innate immunity. Viruses 13(4):560
pubmed: 33810391
pmcid: 8066579
doi: 10.3390/v13040560
Sørensen MD, Sørensen B, GONZALEZ-DOSAL R, Melchjorsen CJ, Weibel J, Wang J, Jun CW, Huanming Y, Kristensen P (2006) Severe Acute Respiratory Syndrome (SARS) Development of Diagnostics and antivirals, vol 1067. Annals of the New York Academy of Sciences, pp 500–505. 1
WHO. COVID-19 (2023) ; https://covid19.who.int/
Costanzo M, De Giglio MA, Roviello GN (2022) Anti-coronavirus vaccines: past investigations on SARS-CoV-1 and MERS-CoV, the approved vaccines from BioNTech/Pfizer, Moderna, Oxford/AstraZeneca and others under Development Against SARSCoV-2 infection. Curr Med Chem 29(1):4–18
pubmed: 34355678
doi: 10.2174/0929867328666210521164809
Rehwinkel JMU, Gack (2020) RIG-I-like receptors: their regulation and roles in RNA sensing. Nat Rev Immunol 20(9):537–551
pubmed: 32203325
pmcid: 7094958
doi: 10.1038/s41577-020-0288-3
Platanias LC (2005) Mechanisms of type-I-and type-II-interferon-mediated signalling. Nat Rev Immunol 5(5):375–386
pubmed: 15864272
doi: 10.1038/nri1604
Rabaan AA, Al-Ahmed SH, Haque S, Sah R, Tiwari R, Malik YS, Dhama K, Yatoo MI, Bonilla-Aldana DK, Rodriguez-Morales AJ (2020) SARS-CoV-2, SARS-CoV, and MERS-COV: a comparative overview. Infez Med 28(2):174–184
pubmed: 32275259
CE Comar, CJ Otter, J Pfannenstiel, E Doerger, DM Renner, LH Tan, S Perlman, NA Cohen, AR Fehr, SR Weiss (2022) MERS-CoV endoribonuclease and accessory proteins jointly evade host innate immunity during infection of lung and nasal epithelial cells. Proc Natl Acad Sci 119(21):e2123208119
Liu DX, Fung TS, Chong KK-L, Shukla A, Hilgenfeld R (2014) Accessory proteins of SARS-CoV and other coronaviruses. Antiviral Res 109:97–109
pubmed: 24995382
pmcid: 7113789
doi: 10.1016/j.antiviral.2014.06.013
Vijay RS, Perlman (2016) Middle East respiratory syndrome and severe acute respiratory syndrome. Curr Opin Virol 16:70–76
pubmed: 26855039
pmcid: 4821769
doi: 10.1016/j.coviro.2016.01.011
Chang C-Y, Liu HM, Chang M-F, Chang SC (2020) Middle East respiratory syndrome coronavirus nucleocapsid protein suppresses type I and type III interferon induction by targeting RIG-I signaling. J Virol 94(13):e00099–e00020
pubmed: 32295922
pmcid: 7307178
doi: 10.1128/JVI.00099-20
Lui P-Y, Wong L-YR, Fung C-L, Siu K-L, Yeung M-L, Yuen K-S, Chan C-P, Woo PC-Y, Yuen K-Y, Jin D-Y (2016) Middle East respiratory syndrome coronavirus M protein suppresses type I interferon expression through the inhibition of TBK1-dependent phosphorylation of IRF3, vol 5. Emerging microbes & infections, pp 1–9. 1
Niemeyer D, Zillinger T, Muth D, Zielecki F, Horvath G, Suliman T, Barchet W, Weber F, Drosten C, Müller MA (2013) Middle East respiratory syndrome coronavirus accessory protein 4a is a type I interferon antagonist. J Virol 87(22):12489–12495
pubmed: 24027320
pmcid: 3807936
doi: 10.1128/JVI.01845-13
Yang Y, Ye F, Zhu N, Wang W, Deng Y, Zhao Z, Tan W (2015) Middle East respiratory syndrome coronavirus ORF4b protein inhibits type I interferon production through both cytoplasmic and nuclear targets. Sci Rep 5(1):1–13
doi: 10.1038/srep17554
Wong L-YR, Ye Z-W, Lui P-Y, Zheng X, Yuan S, Zhu L, Fung S-Y, Yuen K-S, Siu K-L, Yeung M-L (2020) Middle east respiratory syndrome coronavirus ORF8b accessory protein suppresses type I IFN expression by impeding HSP70-dependent activation of IRF3 kinase IKKε. J Immunol 205(6):1564–1579
pubmed: 32747502
doi: 10.4049/jimmunol.1901489
Yang X, Chen X, Bian G, Tu J, Xing Y, Wang Y, Chen Z (2014) Proteolytic processing, deubiquitinase and interferon antagonist activities of Middle East respiratory syndrome coronavirus papain-like protease. J Gen Virol 95(3):614–626
pubmed: 24362959
doi: 10.1099/vir.0.059014-0
Cao D, Duan L, Huang B, Xiong Y, Zhang G, Huang H (2023) The SARS-CoV-2 papain-like protease suppresses type I interferon responses by deubiquitinating STING. Sci Signal 16(783):eadd0082
pubmed: 37130168
doi: 10.1126/scisignal.add0082
Hilgenfeld R (2014) From SARS to MERS: crystallographic studies on coronaviral proteases enable antiviral drug design. FEBS J 281(18):4085–4096
pubmed: 25039866
pmcid: 7163996
doi: 10.1111/febs.12936
He J, Hu L, Huang X, Wang C, Zhang Z, Wang Y, Zhang D, Ye W (2020) Potential of coronavirus 3 C-like protease inhibitors for the development of new anti-SARS-CoV-2 drugs: insights from structures of protease and inhibitors. Int J Antimicrob Agents 56(2):106055
pubmed: 32534187
pmcid: 7286838
doi: 10.1016/j.ijantimicag.2020.106055
Fung S-Y, Siu K-L, Lin H, Yeung ML, Jin D-Y (2021) SARS-CoV-2 main protease suppresses type I interferon production by preventing nuclear translocation of phosphorylated IRF3. Int J Biol Sci 17(6):1547
pubmed: 33907518
pmcid: 8071772
doi: 10.7150/ijbs.59943
Liu Y, Qin C, Rao Y, Ngo C, Feng JJ, Zhao J, Zhang S, Wang T-Y, Carriere J, Savas AC (2021) SARS-CoV-2 Nsp5 demonstrates two distinct mechanisms targeting RIG-I and MAVS to evade the innate immune response. MBio 12(5):e02335–e02321
pubmed: 34544279
pmcid: 8546575
doi: 10.1128/mBio.02335-21
Wu Y, Ma L, Zhuang Z, Cai S, Zhao Z, Zhou L, Zhang J, Wang P-H, Zhao J, Cui J (2020) Main protease of SARS-CoV-2 serves as a bifunctional molecule in restricting type I interferon antiviral signaling. Signal Transduct Target Therapy 5(1):221
doi: 10.1038/s41392-020-00332-2
Fani M, Teimoori A, Ghafari S (2020) Comparison of the COVID-2019 (SARS-CoV-2) pathogenesis with SARS-CoV and MERS-CoV infections. Future Virol 15(5):317–323
doi: 10.2217/fvl-2020-0050
Zhu M, Fang T, Li S, Meng K, Guo D (2015) Bipartite nuclear localization signal controls nuclear import and DNA-binding activity of IFN regulatory factor 3. J Immunol 195(1):289–297
pubmed: 25994966
doi: 10.4049/jimmunol.1500232
Molecular Operating Environment (MOE), version 2022.02; Chemical Computing Group Inc.: Montreal, QC, Canada. (2022) 02; https://www.chemcomp.com/en/Products.htm
De Ioannes P, Escalante CR, Aggarwal AK (2011) Structures of apo IRF-3 and IRF-7 DNA binding domains: effect of loop L1 on DNA binding. Nucleic Acids Res 39(16):7300–7307
pubmed: 21596780
pmcid: 3167601
doi: 10.1093/nar/gkr325
Dampalla CS, Miller MJ, Kim Y, Zabiegala A, Nguyen HN, Madden TK, Thurman HA, Machen AJ, Cooper A, Liu L (2023) Structure-guided design of direct-acting antivirals that exploit the gem-dimethyl effect and potently inhibit 3CL proteases of severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) and middle east respiratory syndrome coronavirus (MERS-CoV). Eur J Med Chem 254:115376
pubmed: 37080108
pmcid: 10105399
doi: 10.1016/j.ejmech.2023.115376
Florio TJ, Lokareddy RK, Yeggoni DP, Sankhala RS, Ott CA, Gillilan RE, Cingolani G (2022) Differential recognition of canonical NF-κB dimers by Importin α3. Nat Commun 13(1):1207
pubmed: 35260573
pmcid: 8904830
doi: 10.1038/s41467-022-28846-z
Pronk S, Páll S, Schulz R, Larsson P, Bjelkmar P, Apostolov R, Shirts MR, Smith JC, Kasson PM, Van Der Spoel D (2013) GROMACS 4.5: a high-throughput and highly parallel open source molecular simulation toolkit. Bioinformatics 29(7):845–854
pubmed: 23407358
pmcid: 3605599
doi: 10.1093/bioinformatics/btt055
Desta IT, Porter KA, Xia B, Kozakov D, Vajda S (2020) Performance and its limits in rigid body protein-protein docking. Structure 28(9):1071–1081e3
pubmed: 32649857
pmcid: 7484347
doi: 10.1016/j.str.2020.06.006
Pierce BG, Wiehe K, Hwang H, Kim B-H, Vreven T, Weng Z (2014) ZDOCK server: interactive docking prediction of protein–protein complexes and symmetric multimers. Bioinformatics 30(12):1771–1773
pubmed: 24532726
pmcid: 4058926
doi: 10.1093/bioinformatics/btu097
Abramson J, Adler J, Dunger J, Evans R, Green T, Pritzel A, Ronneberger O, Willmore L, Ballard AJ, Bambrick J (2024) Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature (630):493–500
Jo S, Kim T, Iyer VG, Im W (2008) CHARMM-GUI: a web‐based graphical user interface for CHARMM. J Comput Chem 29(11):1859–1865
pubmed: 18351591
doi: 10.1002/jcc.20945
Turner P Center For Coastal and Land-Margin Research, Oregon Graduate Institute of Science and Technology; Beaverton, Ore, USA: 2005. XMGRACE, Version. 5: p. 19
Liu Y, Qin C, Rao Y, Ngo C, Feng JJ, Zhao J, Zhang S, Wang T-Y, Carriere J, Savas AC (2021) SARS-CoV-2 Nsp5 demonstrates two distinct mechanisms targeting RIG-I and MAVS to evade the innate immune response. MBio 12(5):02335–02321. https://doi.org/10.1128/mbio
doi: 10.1128/mbio
Zhu X, Fang L, Wang D, Yang Y, Chen J, Ye X, Foda MF, Xiao S (2017) Porcine deltacoronavirus nsp5 inhibits interferon-β production through the cleavage of NEMO. Virology 502:33–38
pubmed: 27984784
doi: 10.1016/j.virol.2016.12.005
Naik NG, Lee S-C, Veronese BH, Ma Z, Toth Z (2022) Interaction of HDAC2 with SARS-CoV-2 NSP5 and IRF3 is not required for NSP5-mediated inhibition of type I interferon signaling pathway. Microbiol Spectr 10(5):e02322–e02322
pubmed: 36173315
pmcid: 9603796
doi: 10.1128/spectrum.02322-22
Tran EJ, King MC, Corbett AH (2014) Macromolecular transport between the nucleus and the cytoplasm: advances in mechanism and emerging links to disease. Biochim et Biophys Acta (BBA)-Molecular Cell Res 1843(11):2784–2795
doi: 10.1016/j.bbamcr.2014.08.003
Ye J, Chen Z, Li Y, Zhao Z, He W, Zohaib A, Song Y, Deng C, Zhang B, Chen H (2017) Japanese encephalitis virus NS5 inhibits type I interferon (IFN) production by blocking the nuclear translocation of IFN regulatory factor 3 and NF-κB. J Virol 91(8). https://doi.org/10.1128/jvi 00039 – 17
Li J, Lu M, Huang B, Lv Y (2018) Porcine circovirus type 2 inhibits interferon-β expression by targeting karyopherin alpha-3 in PK-15 cells. Virology 520:75–82
pubmed: 29793076
doi: 10.1016/j.virol.2018.05.008
Lockbaum GJ, Henes M, Lee JM, Timm J, Nalivaika EA, Thompson PR, Kurt Yilmaz N, Schiffer CA (2021) Pan-3 C protease inhibitor rupintrivir binds SARS-CoV-2 main protease in a unique binding mode. Biochemistry 60(39):2925–2931
pubmed: 34506130
doi: 10.1021/acs.biochem.1c00414
Antonopoulou I, Sapountzaki E, Rova U, Christakopoulos P (2022) Inhibition of the main protease of SARS-CoV-2 (Mpro) by repurposing/designing drug-like substances and utilizing nature’s toolbox of bioactive compounds. Comput Struct Biotechnol J 20:1306–1344
pubmed: 35308802
pmcid: 8920478
doi: 10.1016/j.csbj.2022.03.009
Kelly SM, VanSlyke JK, Musil LS (2007) Regulation of ubiquitin-proteasome system–mediated degradation by Cytosolic Stress. Mol Biol Cell 18(11):4279–4291
pubmed: 17699585
pmcid: 2043544
doi: 10.1091/mbc.e07-05-0487
Fayne D (2013) De-peptidising protein–protein interactions–big jobs for small molecules. Drug Discovery Today: Technol 10(4):e467–e474
doi: 10.1016/j.ddtec.2013.08.002
Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O, Tunyasuvunakool K, Bates R, Žídek A, Potapenko A (2021) Highly accurate protein structure prediction with AlphaFold. Nature 596(7873):583–589
pubmed: 34265844
pmcid: 8371605
doi: 10.1038/s41586-021-03819-2
Gordon DE, Hiatt J, Bouhaddou M, Rezelj VV, Ulferts S, Braberg H, Jureka AS, Obernier K, Guo JZ, Batra J (2020) Comparative host-coronavirus protein interaction networks reveal pan-viral disease mechanisms. Science 370(6521):eabe9403
pubmed: 33060197
pmcid: 7808408
doi: 10.1126/science.abe9403
Messina F, Giombini E, Agrati C, Vairo F, Ascoli Bartoli T, Al Moghazi S, Piacentini M, Locatelli F, Kobinger G, Maeurer M (2020) COVID-19: viral–host interactome analyzed by network based-approach model to study pathogenesis of SARS-CoV-2 infection. J Translational Med 18:1–10
doi: 10.1186/s12967-020-02405-w
Hopfner K-PV, Hornung (2020) Molecular mechanisms and cellular functions of cGAS–STING signalling. Nat Rev Mol Cell Biol 21(9):501–521
pubmed: 32424334
doi: 10.1038/s41580-020-0244-x
Wang W, Zhou Z, Xiao X, Tian Z, Dong X, Wang C, Li L, Ren L, Lei X, Xiang Z (2021) SARS-CoV-2 nsp12 attenuates type I interferon production by inhibiting IRF3 nuclear translocation. Cell Mol Immunol 18(4):945–953
pubmed: 33637958
doi: 10.1038/s41423-020-00619-y
Li W, Qiao J, You Q, Zong S, Peng Q, Liu Y, Hu S, Liu W, Li S, Shu X (2021) SARS-CoV-2 Nsp5 activates NF-κB pathway by upregulating SUMOylation of MAVS. Front Immunol 12:750969
pubmed: 34858407
pmcid: 8631293
doi: 10.3389/fimmu.2021.750969
Li M, Ayyanathan K, Dittmar M, Miller J, Tapescu I, Lee JS, McGrath ME, Xue Y, Vashee S, Schultz DC (2023) SARS-CoV-2 ORF6 protein does not antagonize interferon signaling in respiratory epithelial Calu-3 cells during infection. Mbio, : pp. e01194-23
Lei X, Dong X, Ma R, Wang W, Xiao X, Tian Z, Wang C, Wang Y, Li L, Ren L (2020) Activation and evasion of type I interferon responses by SARS-CoV-2. Nat Commun 11(1):3810
pubmed: 32733001
pmcid: 7392898
doi: 10.1038/s41467-020-17665-9
Liu L, Wei Q, Alvarez X, Wang H, Du Y, Zhu H, Jiang H, Zhou J, Lam P, Zhang L (2011) Epithelial cells lining salivary gland ducts are early target cells of severe acute respiratory syndrome coronavirus infection in the upper respiratory tracts of rhesus macaques. J Virol 85(8):4025–4030
pubmed: 21289121
pmcid: 3126125
doi: 10.1128/JVI.02292-10
Liu Y, Tronser T, Peravali R, Reischl M, Levkin PA (2020) High-throughput screening of cell transfection enhancers using miniaturized droplet microarrays. Adv Biosystems 4(3):1900257
doi: 10.1002/adbi.201900257
Hillyer P, Shepard R, Uehling M, Krenz M, Sheikh F, Thayer KR, Huang L, Yan L, Panda D, Luongo C (2018) Differential responses by human respiratory epithelial cell lines to respiratory syncytial virus reflect distinct patterns of infection control. J Virol 92(15). https://doi.org/10.1128/jvi.02202-17
Xu S, Xie J, Zhang X, Chen L, Bi Y, Li X, Idris A, Feng R (2022) DDX56 antagonizes IFN-β production to enhance EMCV replication by inhibiting IRF3 nuclear translocation. Vet Microbiol 264:109304
pubmed: 34922148
doi: 10.1016/j.vetmic.2021.109304
Hamrashdi MAG, Brady (2022) Regulation of IRF3 activation in human antiviral signaling pathways. Biochem Pharmacol 200:115026
pubmed: 35367198
doi: 10.1016/j.bcp.2022.115026
Hiscott J, Nguyen T, Arguello M, Nakhaei P, Paz S (2006) Manipulation of the nuclear factor-κB pathway and the innate immune response by viruses. Oncogene 25(51):6844–6867
pubmed: 17072332
pmcid: 7100320
doi: 10.1038/sj.onc.1209941
Gottipati K, Holthauzen LMF, Ruggli N, Choi KH (2016) Pestivirus Npro directly interacts with interferon regulatory factor 3 monomer and dimer. J Virol 90(17):7740–7747
pubmed: 27334592
pmcid: 4988160
doi: 10.1128/JVI.00318-16
Cai Z, Zhang M-X, Tang Z, Zhang Q, Ye J, Xiong T-C, Zhang Z-D, Zhong B (2020) USP22 promotes IRF3 nuclear translocation and antiviral responses by deubiquitinating the importin protein KPNA2. J Exp Med, 217(5)
Ma C, Sacco MD, Hurst B, Townsend JA, Hu Y, Szeto T, Zhang X, Tarbet B, Marty MT, Chen Y (2020) Boceprevir, GC-376, and calpain inhibitors II, XII inhibit SARS-CoV-2 viral replication by targeting the viral main protease. Cell Res 30(8):678–692
pubmed: 32541865
pmcid: 7294525
doi: 10.1038/s41422-020-0356-z
Ng CS, Stobart CC, Luo H (2021) Innate immune evasion mediated by picornaviral 3 C protease: possible lessons for coronaviral 3 C-like protease? Rev Med Virol 31(5):1–22
pubmed: 33624382
doi: 10.1002/rmv.2206
Hu Y, Ma C, Szeto T, Hurst B, Tarbet B, Wang J (2021) Boceprevir, calpain inhibitors II and XII, and GC-376 have broad-spectrum antiviral activity against coronaviruses. ACS Infect Dis 7(3):586–597
pubmed: 33645977
doi: 10.1021/acsinfecdis.0c00761
Wang D, Fang L, Shi Y, Zhang H, Gao L, Peng G, Chen H, Li K, Xiao S (2016) Porcine epidemic diarrhea virus 3 C-like protease regulates its interferon antagonism by cleaving NEMO. J Virol 90(4):2090–2101
pubmed: 26656704
pmcid: 4733996
doi: 10.1128/JVI.02514-15
Masters JR (2002) HeLa cells 50 years on: the good, the bad and the ugly. Nat Rev Cancer 2(4):315–319
pubmed: 12001993
doi: 10.1038/nrc775
Sharma SV, Haber DA, Settleman J (2010) Cell line-based platforms to evaluate the therapeutic efficacy of candidate anticancer agents. Nat Rev Cancer 10(4):241–253
pubmed: 20300105
doi: 10.1038/nrc2820
Kube D, Sontich U, Fletcher D, Davis PB (2001) Proinflammatory cytokine responses to P. Aeruginosa infection in human airway epithelial cell lines. Am J Physiology-Lung Cell Mol Physiol 280(3):L493–L502
doi: 10.1152/ajplung.2001.280.3.L493
Zhou M, Li Q, Wang R (2016) Current experimental methods for characterizing protein–protein interactions. ChemMedChem 11(8):738–756
pubmed: 26864455
pmcid: 7162211
doi: 10.1002/cmdc.201500495
Arnoldo A, Kittanakom S, Heisler LE, Mak AB, Shukalyuk AI, Torti D, Moffat J, Giaever G, Nislow C (2014) A genome scale overexpression screen to reveal drug activity in human cells. Genome Med 6:1–16
doi: 10.1186/gm549
Skariyachan S, Challapilli SB, Packirisamy S, Kumargowda ST, Sridhar VS (2019) Recent aspects on the pathogenesis mechanism, animal models and novel therapeutic interventions for Middle East respiratory syndrome coronavirus infections. Front Microbiol 10:569
pubmed: 30984127
pmcid: 6448012
doi: 10.3389/fmicb.2019.00569