دورية أكاديمية
Viroporin-like activity of the hairpin transmembrane domain of African swine fever virus B169L protein.
| العنوان: | Viroporin-like activity of the hairpin transmembrane domain of African swine fever virus B169L protein. |
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| المؤلفون: | Gladue DP; Plum Island Animal Disease Center, ARS, USDA, Greenport, New York, USA., Gomez-Lucas L; Instituto Biofisika (CSIC-UPV/EHU) and Department of Biochemistry and Molecular Biology, University of the Basque Country, Bilbao, Spain., Largo E; Department of Immunology, Microbiology and Parasitology, Faculty of Medicine and Nursing, University of the Basque Country (UPV/EHU), Leioa, Spain., Ramirez-Medina E; Plum Island Animal Disease Center, ARS, USDA, Greenport, New York, USA., Torralba J; Instituto Biofisika (CSIC-UPV/EHU) and Department of Biochemistry and Molecular Biology, University of the Basque Country, Bilbao, Spain., Queralt-Martín M; Laboratory of Molecular Biophysics. Department of Physics, University Jaume I, Castello, Castellón, Spain., Alcaraz A; Laboratory of Molecular Biophysics. Department of Physics, University Jaume I, Castello, Castellón, Spain., Velazquez-Salinas L; Plum Island Animal Disease Center, ARS, USDA, Greenport, New York, USA., Nieva JL; Instituto Biofisika (CSIC-UPV/EHU) and Department of Biochemistry and Molecular Biology, University of the Basque Country, Bilbao, Spain., Borca MV; Plum Island Animal Disease Center, ARS, USDA, Greenport, New York, USA. |
| المصدر: | Journal of virology [J Virol] 2024 Aug 20; Vol. 98 (8), pp. e0023124. Date of Electronic Publication: 2024 Jul 09. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society For Microbiology Country of Publication: United States NLM ID: 0113724 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5514 (Electronic) Linking ISSN: 0022538X NLM ISO Abbreviation: J Virol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Washington Dc : American Society For Microbiology Original Publication: Baltimore, American Society for Microbiology. |
| مواضيع طبية MeSH: | African Swine Fever Virus*/genetics , African Swine Fever Virus*/metabolism, Animals ; Swine ; Endoplasmic Reticulum/metabolism ; Endoplasmic Reticulum/virology ; Protein Domains ; Viroporin Proteins/metabolism ; Viroporin Proteins/genetics ; African Swine Fever/virology ; African Swine Fever/metabolism ; Viral Proteins/metabolism ; Viral Proteins/genetics ; Viral Proteins/chemistry ; Amino Acid Sequence |
| مستخلص: | African swine fever virus (ASFV) is the causative agent of a contagious disease affecting wild and domestic swine. The function of B169L protein, as a potential integral structural membrane protein, remains to be experimentally characterized. Using state-of-the-art bioinformatics tools, we confirm here earlier predictions indicating the presence of an integral membrane helical hairpin, and further suggest anchoring of this protein to the ER membrane, with both terminal ends facing the lumen of the organelle. Our evolutionary analysis confirmed the importance of purifying selection in the preservation of the identified domains during the evolution of B169L in nature. Also, we address the possible function of this hairpin transmembrane domain (HTMD) as a class IIA viroporin. Expression of GFP fusion proteins in the absence of a signal peptide supported B169L insertion into the ER as a Type III membrane protein and the formation of oligomers therein. Overlapping peptides that spanned the B169L HTMD were reconstituted into ER-like membranes and the adopted structures analyzed by infrared spectroscopy. Consistent with the predictions, B169L transmembrane sequences adopted α-helical conformations in lipid bilayers. Moreover, single vesicle permeability assays demonstrated the assembly of lytic pores in ER-like membranes by B169L transmembrane helices, a capacity confirmed by ion-channel activity measurements in planar bilayers. Emphasizing the relevance of these observations, pore-forming activities were not observed in the case of transmembrane helices derived from EP84R, another ASFV protein predicted to anchor to membranes through a α-helical HTMD. Overall, our results support predictions of viroporin-like function for the B169L HTMD.IMPORTANCEAfrican swine fever (ASF), a devastating disease affecting domestic swine, is widely spread in Eurasia, producing significant economic problems in the pork industry. Approaches to prevent/cure the disease are mainly restricted to the limited information concerning the role of most of the genes encoded by the large (160-170 kba) virus genome. In this report, we present the experimental data on the functional characterization of the African swine fever virus (ASFV) gene B169L. Data presented here indicates that the B169L gene encodes for an essential membrane-associated protein with a viroporin function. Competing Interests: The authors declare no conflict of interest. |
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| معلومات مُعتمدة: | IT1449-22 Basque Government |
| فهرسة مساهمة: | Keywords: ASF; ASFV; African swine fever; African swine fever virus; B169L; viroporin |
| المشرفين على المادة: | 0 (Viroporin Proteins) 0 (Viral Proteins) |
| تواريخ الأحداث: | Date Created: 20240709 Date Completed: 20240820 Latest Revision: 20240822 |
| رمز التحديث: | 20240822 |
| مُعرف محوري في PubMed: | PMC11334534 |
| DOI: | 10.1128/jvi.00231-24 |
| PMID: | 38980063 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1098-5514 |
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| DOI: | 10.1128/jvi.00231-24 |