An Unbiased Approach to Mapping the Signaling Network of the Pseudorabies Virus US3 Protein

Overview

Journal Pathogens

Date 2020 Nov 10

PMID 33167340

Citations 5

Authors

Robert J J Jansens

Sandra Marmiroli

Herman W Favoreel

Affiliations

Soon will be listed here.

Abstract

The US3 serine/threonine protein kinase is conserved among the alphaherpesvirus family and represents an important virulence factor. US3 plays a role in viral nuclear egress, induces dramatic alterations of the cytoskeleton, represses apoptosis, enhances gene expression and modulates the immune response. Although several substrates of US3 have been identified, an unbiased screen to identify US3 phosphorylation targets has not yet been described. Here, we perform a shotgun and phosphoproteomics analysis of cells expressing the US3 protein of pseudorabies virus (PRV) to identify US3 phosphorylation targets in an unbiased way. We identified several cellular proteins that are differentially phosphorylated upon US3 expression and validated the phosphorylation of lamin A/C at serine 404, both in US3-transfected and PRV-infected cells. These results provide new insights into the signaling network of the US3 protein kinase and may serve as a basis for future research into the role of the US3 protein in the viral replication cycle.

Citing Articles

Alphaherpesvirus-mediated remodeling of the cellular transcriptome results in depletion of m6A-containing transcripts.

Jansens R, Olarerin-George A, Verhamme R, Mirza A, Jaffrey S, Favoreel H iScience. 2023; 26(8):107310.

PMID: 37575180 PMC: 10415716. DOI: 10.1016/j.isci.2023.107310.

Guanylate-binding protein 1 inhibits nuclear delivery of pseudorabies virus by disrupting structure of actin filaments.

Zhang X, Du Q, Chen G, Jiang Y, Huang K, Li L Vet Res. 2023; 54(1):21.

PMID: 36918936 PMC: 10015811. DOI: 10.1186/s13567-023-01154-0.

The Deletion of US3 Gene of Pseudorabies Virus (PRV) ΔgE/TK Strain Induces Increased Immunogenicity in Mice.

Deng M, Sun Y, Ding C, Xu X, Guo Z, Han Z Vaccines (Basel). 2022; 10(10).

PMID: 36298468 PMC: 9612271. DOI: 10.3390/vaccines10101603.

Alphaherpesvirus US3 protein-mediated inhibition of the m6A mRNA methyltransferase complex.

Jansens R, Verhamme R, Mirza A, Olarerin-George A, Van Waesberghe C, Jaffrey S Cell Rep. 2022; 40(3):111107.

PMID: 35858564 PMC: 9347262. DOI: 10.1016/j.celrep.2022.111107.

Pseudorabies Virus Infections.

Klupp B Pathogens. 2021; 10(6).

PMID: 34201049 PMC: 8226705. DOI: 10.3390/pathogens10060719.

References

de Wind N, Domen J, Berns A . Herpesviruses encode an unusual protein-serine/threonine kinase which is nonessential for growth in cultured cells. J Virol. 1992; 66(9):5200-9. PMC: 289072. DOI: 10.1128/JVI.66.9.5200-5209.1992. View

Chuluunbaatar U, Mohr I . A herpesvirus kinase that masquerades as Akt: you don't have to look like Akt, to act like it. Cell Cycle. 2011; 10(13):2064-8. PMC: 3154360. DOI: 10.4161/cc.10.13.16242. View

Bertacchini J, Beretti F, Cenni V, Guida M, Gibellini F, Mediani L . The protein kinase Akt/PKB regulates both prelamin A degradation and Lmna gene expression. FASEB J. 2013; 27(6):2145-55. DOI: 10.1096/fj.12-218214. View

Miranda L, Carpentier S, Platek A, Hussain N, Gueuning M, Vertommen D . AMP-activated protein kinase induces actin cytoskeleton reorganization in epithelial cells. Biochem Biophys Res Commun. 2010; 396(3):656-61. DOI: 10.1016/j.bbrc.2010.04.151. View

Rontogianni S, Iskit S, van Doorn S, Peeper D, Altelaar M . Combined EGFR and ROCK Inhibition in Triple-negative Breast Cancer Leads to Cell Death Via Impaired Autophagic Flux. Mol Cell Proteomics. 2019; 19(2):261-277. PMC: 7000121. DOI: 10.1074/mcp.RA119.001800. View

Perez-Riverol Y, Csordas A, Bai J, Bernal-Llinares M, Hewapathirana S, Kundu D . The PRIDE database and related tools and resources in 2019: improving support for quantification data. Nucleic Acids Res. 2018; 47(D1):D442-D450. PMC: 6323896. DOI: 10.1093/nar/gky1106. View

Jacob T, Van den Broeke C, Van Troys M, Waterschoot D, Ampe C, Favoreel H . Alphaherpesviral US3 kinase induces cofilin dephosphorylation to reorganize the actin cytoskeleton. J Virol. 2013; 87(7):4121-6. PMC: 3624233. DOI: 10.1128/JVI.03107-12. View

de Wind N, Zijderveld A, Glazenburg K, Gielkens A, Berns A . Linker insertion mutagenesis of herpesviruses: inactivation of single genes within the Us region of pseudorabies virus. J Virol. 1990; 64(10):4691-6. PMC: 247954. DOI: 10.1128/JVI.64.10.4691-4696.1990. View

Poon A, Gu H, Roizman B . ICP0 and the US3 protein kinase of herpes simplex virus 1 independently block histone deacetylation to enable gene expression. Proc Natl Acad Sci U S A. 2006; 103(26):9993-8. PMC: 1502567. DOI: 10.1073/pnas.0604142103. View

10.

Geenen K, Favoreel H, Olsen L, Enquist L, Nauwynck H . The pseudorabies virus US3 protein kinase possesses anti-apoptotic activity that protects cells from apoptosis during infection and after treatment with sorbitol or staurosporine. Virology. 2004; 331(1):144-50. DOI: 10.1016/j.virol.2004.10.027. View

11.

Olsen L, Chng T, Card J, Enquist L . Role of pseudorabies virus Us3 protein kinase during neuronal infection. J Virol. 2006; 80(13):6387-98. PMC: 1488934. DOI: 10.1128/JVI.00352-06. View

12.

Benetti L, Roizman B . Herpes simplex virus protein kinase US3 activates and functionally overlaps protein kinase A to block apoptosis. Proc Natl Acad Sci U S A. 2004; 101(25):9411-6. PMC: 438990. DOI: 10.1073/pnas.0403160101. View

13.

Kulej K, Avgousti D, Sidoli S, Herrmann C, Della Fera A, Kim E . Time-resolved Global and Chromatin Proteomics during Herpes Simplex Virus Type 1 (HSV-1) Infection. Mol Cell Proteomics. 2017; 16(4 suppl 1):S92-S107. PMC: 5393384. DOI: 10.1074/mcp.M116.065987. View

14.

Kimman T, de Wind N, Pol J, Berns A, Gielkens A . Contribution of single genes within the unique short region of Aujeszky's disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity. J Gen Virol. 1992; 73 ( Pt 2):243-51. DOI: 10.1099/0022-1317-73-2-243. View

15.

Trivedi N, Marsh P, Goold R, Wood-Kaczmar A, Gordon-Weeks P . Glycogen synthase kinase-3beta phosphorylation of MAP1B at Ser1260 and Thr1265 is spatially restricted to growing axons. J Cell Sci. 2005; 118(Pt 5):993-1005. DOI: 10.1242/jcs.01697. View

16.

Chuluunbaatar U, Roller R, Feldman M, Brown S, Shokat K, Mohr I . Constitutive mTORC1 activation by a herpesvirus Akt surrogate stimulates mRNA translation and viral replication. Genes Dev. 2010; 24(23):2627-39. PMC: 2994037. DOI: 10.1101/gad.1978310. View

17.

Walters M, Kinchington P, Banfield B, Silverstein S . Hyperphosphorylation of histone deacetylase 2 by alphaherpesvirus US3 kinases. J Virol. 2010; 84(19):9666-76. PMC: 2937806. DOI: 10.1128/JVI.00981-10. View

18.

Wang S, Wang K, Lin R, Zheng C . Herpes simplex virus 1 serine/threonine kinase US3 hyperphosphorylates IRF3 and inhibits beta interferon production. J Virol. 2013; 87(23):12814-27. PMC: 3838156. DOI: 10.1128/JVI.02355-13. View

19.

Rosok O, Pedeutour F, Ree A, Aasheim H . Identification and characterization of TESK2, a novel member of the LIMK/TESK family of protein kinases, predominantly expressed in testis. Genomics. 1999; 61(1):44-54. DOI: 10.1006/geno.1999.5922. View

20.

Mendes Maia T, Staes A, Plasman K, Pauwels J, Boucher K, Argentini A . Simple Peptide Quantification Approach for MS-Based Proteomics Quality Control. ACS Omega. 2020; 5(12):6754-6762. PMC: 7114614. DOI: 10.1021/acsomega.0c00080. View