Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

Chuntian Li; Yuncheng Li; Ranqing Cheng; Miaomiao Li; Mudan Zhang; Zhiyuan Zhu; Ping Yang; Qinxue Hu; Yalan Liu

doi:10.1016/j.virs.2025.08.004

October 2025

Citation: Chuntian Li, Yuncheng Li, Ranqing Cheng, Miaomiao Li, Mudan Zhang, Zhiyuan Zhu, Ping Yang, Qinxue Hu, Yalan Liu. Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway .VIROLOGICA SINICA, 2025, 40(5) : 778-792. http://dx.doi.org/10.1016/j.virs.2025.08.004

Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

Chuntian Li ^a ,
Yuncheng Li ^b,c ,
Ranqing Cheng ^b,c ,
Miaomiao Li ^b,c ,
Mudan Zhang ^b ,
Zhiyuan Zhu ^b,c ,
Ping Yang ^b ,
Qinxue Hu ^{b,c
,,} ,
Yalan Liu ^{b,c,d
,,}

a. Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Henan University, School of Medicine, Henan University, Kaifeng, 475004, China;
b. State Key Laboratory of Virology and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China;
c. Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China;
d. Hubei Jiangxia Laboratory, Wuhan, 430200, China

Corresponding author: Qinxue Hu, qhu@wh.iov.cn
Yalan Liu, liuyl@wh.iov.cn
Received Date: 28 April 2025
Accepted Date: 27 August 2025
Available online: 02 September 2025

Abstract

Herpes simplex virus 2 (HSV-2) is a major pathogen causing neonatal herpes and increasing the risk of human immunodeficiency virus 1 (HIV-1) infection. However, the mechanisms underlying host restriction of HSV-2 infection are still not fully understood. The ubiquitously expressed transcript isoform 2 (UXT-V2), an α-type prefoldin protein, functions as a versatile transcription factor associated with numerous human tumors, but its role in viral infection remains unclear. In this study, we found that ectopic expression of UXT-V2 significantly inhibited HSV-2 replication, while knockout of endogenously expressed UXT-V2 promoted HSV-2 proliferation. Further analysis revealed that UXT-V2 restricts HSV-2 replication independent of its role in regulating NF-κB. In the context of HSV-2 infection or in viral glycoprotein B (gB)-transfected cells, UXT-V2 facilitates K48-linked ubiquitination of gB, leading to its degradation via the proteasome pathway, thereby inhibiting viral replication. Furthermore, we identified that UXT-V2 interacts with gB, recruiting the E3 ligase TRIM21 to facilitate K48-linked ubiquitination of gB. HSV-2, in turn, reduces the abundance of UXT-V2 proteins both in vitro and in mice, highlighting the complexity of HSV-2-host interactions. Collectively, our findings, for the first time, demonstrate an anti-HSV-2 role of UXT-V2, unveiling a novel host immune defense mechanism involved in regulating glycoprotein homeostasis.
- Herpes simplex virus 2 (HSV-2)
- , UXT-V2
- , Glycoprotein B (gB)
- , Ubiquitination

Electronic Supplementary Material

10.1016j.virs.2025.08.004-ESM2.xlsx
10.1016j.virs.2025.08.004-ESM1.docx
10.1016j.virs.2025.08.004-ESM3.xlsx

References
1. Abe, A., Takahashi-Niki, K., Takekoshi, Y., Shimizu, T., Kitaura, H., Maita, H., Iguchi-Ariga, S.M., Ariga, H., 2013. Prefoldin plays a role as a clearance factor in preventing proteasome inhibitor-induced protein aggregation. J Biol Chem, 288, 27764-27776.
2. Bender, F.C., Whitbeck, J.C., Ponce De Leon, M., Lou, H., Eisenberg, R.J., Cohen, G.H., 2003. Specific Association of Glycoprotein B with Lipid Rafts during Herpes Simplex Virus Entry. J Virol, 77, 9542-9552.
3. Byrne, C.M., Gantt, S., Coombs, D., 2018. Effects of spatiotemporal HSV-2 lesion dynamics and antiviral treatment on the risk of HIV-1 acquisition. PLoS Comput Biol, 14, e1006129.
4. Calistri, A., Sette, P., Salata, C., Cancellotti, E., Forghieri, C., Comin, A., GoTtlinger, H., Campadelli-Fiume, G., Palu, G., Parolin, C., 2007. Intracellular Trafficking and Maturation of Herpes Simplex Virus Type 1 gB and Virus Egress Require Functional Biogenesis of Multivesicular Bodies. J Virol, 81, 11468-11478.
5. Chang, L.S., Wang, J.T., Doong, S.L., Lee, C.P., Chang, C.W., Tsai, C.H., Yeh, S.W., Hsieh, C.Y., Chen, M.R., 2012. Epstein-Barr virus BGLF4 kinase downregulates NF-kappaB transactivation through phosphorylation of coactivator UXT. J Virol, 86, 12176-12186.
6. Chen, S., Chen, K., Zhang, Q., Cheng, H., Zhou, R., 2013. Regulation of the transcriptional activation of the androgen receptor by the UXT-binding protein VHL. Biochem J, 456, 55-66.
7. Cheshenko, N., Herold, B.C., 2002. Glycoprotein B plays a predominant role in mediating herpes simplex virus type 2 attachment and is required for entry and cell-to-cell spread. J Gen Virol, 83, 2247-2255.
8. Cooper, R.S., Heldwein, E.E., 2015. Herpesvirus gB: A Finely Tuned Fusion Machine. Viruses, 7, 6552-6569.
9. Crameri, M., Bauer, M., Caduff, N., Walker, R., Steiner, F., Franzoso, F.D., Gujer, C., Boucke, K., Kucera, T., Zbinden, A., Munz, C., Fraefel, C., Greber, U.F., Pavlovic, J., 2018. MxB is an interferon-induced restriction factor of human herpesviruses. Nat Commun, 9, 1980.
10. El Kasmi, I., Khadivjam, B., Lackman, M., Duron, J., Bonneil, E., Thibault, P., Lippe, R., Sandri-Goldin, R.M., 2018. Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion. J Virol, 92, e01281-17.
11. Enunlu, I., Ozansoy, M., Basak, A.N., 2011. Alfa-class prefoldin protein UXT is a novel interacting partner of Amyotrophic Lateral Sclerosis 2 (Als2) protein. Biochem Biophys Res Commun, 413, 471-475.
12. Everett, R.D., Johnson, K.E., Bottero, V., Flaherty, S., Dutta, S., Singh, V.V., Chandran, B., 2014. IFI16 Restricts HSV-1 Replication by Accumulating on the HSV-1 Genome, Repressing HSV-1 Gene Expression, and Directly or Indirectly Modulating Histone Modifications. PLoS Pathogens, 10, e1004503.
13. Fan, Z., Grantham, M.L., Smith, M.S., Anderson, E.S., Cardelli, J.A., Muggeridge, M.I., 2002. Truncation of Herpes Simplex Virus Type 2 Glycoprotein B Increases Its Cell Surface Expression and Activity in Cell-Cell Fusion, but These Properties Are Unrelated. J Virol, 76, 9271-9283.
14. Groves, M.J., 2016. Genital Herpes: A Review. Am Fam Physician, 93, 928-934.
15. Guan, X., Zhang, M., Fu, M., Luo, S., Hu, Q., 2019. Herpes Simplex Virus Type 2 Immediate Early Protein ICP27 Inhibits IFN-beta Production in Mucosal Epithelial Cells by Antagonizing IRF3 Activation. Front Immunol, 10, 290.
16. Hensler, H.R., Tomaszewski, M.J., Rappocciolo, G., Rinaldo, C.R., Jenkins, F.J., 2014. Human herpesvirus 8 glycoprotein B binds the entry receptor DC-SIGN. Virus Res, 190, 97-103.
17. Huang, Y., Chen, L., Zhou, Y., Liu, H., Yang, J., Liu, Z.,Wang, C., 2011. UXT-V1 protects cells against TNF-induced apoptosis through modulating complex II formation. Mol Biol Cell, 22, 1389-1397.
18. Huang, Y., Liu, H., Ge, R., Zhou, Y., Lou, X., Wang, C., 2012. UXT-V1 facilitates the formation of MAVS antiviral signalosome on mitochondria. J Immunol, 188, 358-366.
19. Huehn, J., Beyer, M., 2015. Epigenetic and transcriptional control of Foxp3+ regulatory T cells. Semin Immunol, 27, 10-18.
20. James, C., Harfouche, M., Welton, N.J., Turner, K.M., Abu-Raddad, L.J., Gottlieb, S.L., Looker, K.J., 2020. Herpes simplex virus: global infection prevalence and incidence estimates, 2016. Bull World Health Organ., 98:315-329.
21. Jenssen, H., Sandvik, K., Andersen, J.H., Hancock, R.E., Gutteberg, T.J., 2008. Inhibition of HSV cell-to-cell spread by lactoferrin and lactoferricin. Antiviral Res, 79, 192-198.
22. Kimura, Y., Nagao, A., Fujioka, Y., Satou, A., Taira, T., Iguchi-Ariga, S.M., Ariga, H., 2007. MM-1 facilitates degradation of c-Myc by recruiting proteasome and a novel ubiquitin E3 ligase. Int J Oncol, 31, 829-836.
23. Li, C., Guan, X., Du, T., Jin, W., Wu, B., Liu, Y., Wang, P., Hu, B., Griffin, G.E., Shattock, R.J., Hu, Q., 2015. Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9. J Gen Virol, 96, 2381-2393.
24. Li, H., Gao, Q., Shao, Y., Sun, B., Wang, F., Qiao, Y., Wang, N., Liu, S., 2018. Gallid Herpesvirus 1 Initiates Apoptosis in Uninfected Cells through Paracrine Repression of p53. Journal of Virology, 92.
25. Li, W., Wang, L., Jiang, C., Li, H., Zhang, K., Xu, Y., Hao, Q., Li, M., Xue, X., Qin, X., Zhang, C., Wang, H., Zhang, W., Zhang, Y., 2014. UXT is a novel regulatory factor of regulatory T cells associated with Foxp3. Eur J Immunol, 44, 533-544.
26. Liu, Y., Zhu, Z., Li, Y., Yang, M., Hu, Q., 2023. Migrasomes released by HSV-2-infected cells serve as a conveyance for virus spread. Virol Sin. 38:643-645.
27. Luo, S., Hu, K., He, S., Wang, P., Zhang, M., Huang, X., Du, T., Zheng, C., Liu, Y., Hu, Q., 2015. Contribution of N-linked glycans on HSV-2 gB to cell-cell fusion and viral entry. Virology, 483, 72-82.
28. Markus, S.M., Taneja, S.S., Logan, S.K., Li, W., Ha, S., Hittelman, A.B., Rogatsky, I., Garabedian, M.J., 2002. Identification and characterization of ART-27, a novel coactivator for the androgen receptor N terminus. Mol Biol Cell, 13, 670-682.
29. Mulvey, M., Arias, C., Mohr, I., 2007. Maintenance of endoplasmic reticulum (ER) homeostasis in herpes simplex virus type 1-infected cells through the association of a viral glycoprotein with PERK, a cellular ER stress sensor. J Virol, 81, 3377-3390.
30. Pan, M., Yin, Y., Wang, X., Wang, Q., Zhang, L., Hu, H., Wang, C., 2020. Mice deficient in UXT exhibit retinitis pigmentosa-like features via aberrant autophagy activation. Autophagy, 17, 1873-1888.
31. Rao, P., Pham, H.T., Kulkarni, A., Yang, Y., Liu, X., Knipe, D.M., Cresswell, P., Yuan, W., 2011. Herpes Simplex Virus 1 Glycoprotein B and US3 Collaborate To Inhibit CD1d Antigen Presentation and NKT Cell Function. J Virol, 85, 8093-8104.
32. Sanchez-Morgan, N., Kirsch, K.H., Trackman, P.C., Sonenshein, G.E., 2017. UXT Is a LOX-PP Interacting Protein That Modulates Estrogen Receptor Alpha Activity in Breast Cancer Cells. J Cellular Biochem, 118, 2347-2356.
33. Schroer, A., Schneider, S., Ropers, H., Nothwang, H., 1999. Cloning and characterization of UXT, a novel gene in human Xp11, which is widely and abundantly expressed in tumor tissue. Genomics, 56, 340-343.
34. Sethurathinam, S., Singh, L.P., Panneerselvam, P., Byrne, B., Ding, J.L., 2013. UXT plays dual opposing roles on SARM-induced apoptosis. FEBS Lett, 587, 3296-3302.
35. Sparrer, K.M.J., Gableske, S., Zurenski, M.A., Parker, Z.M., Full, F., Baumgart, G.J., Kato, J., Pacheco-Rodriguez, G., Liang, C., Pornillos, O., Moss, J., Vaughan, M., Gack, M.U., 2017. TRIM23 mediates virus-induced autophagy via activation of TBK1. Nat Microbiol, 2, 1543-1557.
36. Stampfer, S.D., Lou, H., Cohen, G.H., Eisenberg, R.J., Heldwein, E.E., 2010. Structural basis of local, pH-dependent conformational changes in glycoprotein B from herpes simplex virus type 1. J Virol, 84, 12924-12933.
37. Su, S.K., Li, C.Y., Lei, P.J., Wang, X., Zhao, Q.Y., Cai, Y., Wang, Z., Li, L., Wu, M., 2016. The EZH1-SUZ12 complex positively regulates the transcription of NF-kappaB target genes through interaction with UXT. J Cell Sci, 129, 2343-2353.
38. Suazo, P.A., Tognarelli, E.I., Kalergis, A.M., Gonzalez, P.A., 2015. Herpes simplex virus 2 infection: molecular association with HIV and novel microbicides to prevent disease. Med Microbiol Immunol, 204, 161-176.
39. Sun, S., Tang, Y., Lou, X., Zhu, L., Yang, K., Zhang, B., Shi, H., Wang, C., 2007a. UXT is a novel and essential cofactor in the NF-kappaB transcriptional enhanceosome. J Cell Biol, 178, 231-244.
40. Tan, Z., Zhang, W., Sun, J., Fu, Z., Ke, X., Zheng, C., Zhang, Y., Li, P., Liu, Y., Hu, Q., Wang, H., Zheng, Z., 2018. ZIKV infection activates the IRE1-XBP1 and ATF6 pathways of unfolded protein response in neural cells. J Neuroinflammation, 15, 275.
41. Taneja, S.S., Ha, S., Swenson, N.K., Torra, I.P., Rome, S., Walden, P.D., Huang, H.Y., Shapiro, E., Garabedian, M.J., Logan, S.K., 2004. ART-27, an androgen receptor coactivator regulated in prostate development and cancer. J Biol Chem, 279, 13944-13952.
42. Temme, S., Eis-Hubinger, A.M., Mclellan, A.D., Koch, N., 2010. The herpes simplex virus-1 encoded glycoprotein B diverts HLA-DR into the exosome pathway. J Immunol, 184, 236-243.
43. Thomas, P.A., Mita, P., Ha, S.,Logan, S.K., 2018. Role of the Unconventional Prefoldin Proteins URI and UXT in Transcription Regulation. Adv Exp Med Biol, 1106, 85-94.
44. Thanos, D., Maniatis, T., 1995. Virus induction of human ifnβ gene expression requires the assembly of an enhanceosome. Cell, 83: 1091-1100.
45. Wang, C.Y., Hour, M.J., Lai, H.C., Chen, C.H., Chang, P.J., Huang, S.H., Lin, C.W., 2018. Epigallocatechin-3-gallate inhibits the early stages of Japanese encephalitis virus infection. Virus Res, 253, 140-146.
46. Wang, D., Fang, L., Li, K., Zhong, H., Fan, J., Ouyang, C., Zhang, H., Duan, E., Luo, R., Zhang, Z., Liu, X., Chen, H., Xiao, S., 2012. Foot-and-mouth disease virus 3C protease cleaves NEMO to impair innate immune signaling. J Virol, 86, 9311-9322.
47. Wang, S., Chi, X., Wei, H., Chen, Y., Chen, Z., Huang, S., Chen, J.L., 2014. Influenza A virus-induced degradation of eukaryotic translation initiation factor 4B contributes to viral replication by suppressing IFITM3 protein expression. J Virol, 88, 8375-8385.
48. Weed, D.J., Dollery, S.J., Komala Sari, T., Nicola, A.V., 2018. Acidic pH Mediates Changes in Antigenic and Oligomeric Conformation of Herpes Simplex Virus gB and Is a Determinant of Cell-Specific Entry. J Virol, 92, e01034-01018.
49. Wright, C.C., Wisner, T.W., Hannah, B.P., Eisenberg, R.J., Cohen, G.H., Johnson, D.C., 2009. Fusion between perinuclear virions and the outer nuclear membrane requires the fusogenic activity of herpes simplex virus gB. J Virol, 83, 11847-11856.
50. Xiao, Y., Ma, Z., Wang, R., Yang, L., Nan, Y., Zhang, Y.J., 2016. Downregulation of protein kinase PKR activation by porcine reproductive and respiratory syndrome virus at its early stage infection. Vet Microbiol, 187, 1-7.
51. Yang, J., Zhao, Y.L., Wu, Z.Q., Si, Y.L., Meng, Y.G., Fu, X.B., Mu, Y.M., Han, W.D., 2009. The single-macro domain protein LRP16 is an essential cofactor of androgen receptor. Endocr Relat Cancer, 16, 139-153.
52. Yoon, M.J., Choi, B., Kim, E.J., Ohk, J., Yang, C., Choi, Y.G., Lee, J., Kang, C., Song, H.K., Kim, Y.K., Woo, J.-S., Cho, Y., Choi, E.-J., Jung, H., Kim, C., 2021. UXT chaperone prevents proteotoxicity by acting as an autophagy adaptor for p62-dependent aggrephagy. Nat Commun, 12, 1955.
53. Zeng, J., Xiang, W., Zhang, Y., Huang, C., Chen, K., Chen, Z., 2019. Ubiquitous expressed transcript promotes tumorigenesis by acting as a positive modulator of the polycomb repressive complex 2 in clear cell renal cell carcinoma. BMC Cancer, 19, 874.
54. Zhang, M., Liu, Y., Wang, P., Guan, X., He, S., Luo, S., Li, C., Hu, K., Jin, W., Du, T., Yan, Y., Zhang, Z., Zheng, Z., Wang, H., Hu, Q., 2015. HSV-2 immediate-early protein US1 inhibits IFN-beta production by suppressing association of IRF-3 with IFN-beta promoter. J Immunol, 194, 3102-3115.
55. Zhang, M., Zhang, M.-X., Zhang, Q., Zhu, G.F., Yuan, L., Zhang, D.-E., Zhu, Q., Yao, J., Shu, H.B., Zhong, B., 2016. USP18 recruits USP20 to promote innate antiviral response through deubiquitinating STING/MITA. Cell Res, 26, 1302-1319.
56. Zhao, H., Wang, Q., Zhang, H., Liu, Q., Du, X., Richter, M., Greene, M.I., 2005. UXT is a novel centrosomal protein essential for cell viability. Mol Biol Cell, 16, 5857-5865.
Proportional views