Suwipa RamphanSarawut KhongwichitChonticha SaisawangDuangnapa KovanichAlbert J. KettermanSukathida UbolPrasert AuewarakulSittiruk RoytrakulDuncan R. SmithAtichat KuadkitkanMahidol UniversityThailand National Center for Genetic Engineering and BiotechnologyFaculty of Medicine, Siriraj Hospital, Mahidol University2019-08-232019-08-232018-07-01Proteomics - Clinical Applications. Vol.12, No.4 (2018)18628354186283462-s2.0-85034071599https://repository.li.mahidol.ac.th/handle/20.500.14594/45118© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim Purpose: Chikungunya virus (CHIKV) is a mosquito transmitted alphavirus that causes chikungunya fever in humans. The CHIKV non-structural protein 2 (nsP2) is a multifunctional protein that additionally modulates the host cell to dampen the innate immune response and inhibit other cellular processes. Experimental design: To further investigate the interactions of nsP2 with host cells, the protease domain of CHIKV nsP2 (nsP2-pro) is transfected into Hela cells, and differential protein expression is detected by 2D polyacrylamide gel electrophoresis. Results: A total of 21 differentially regulated (six upregulated, 15 downregulated) spots are observed, of which five are identified by mass spectrometry. The downregulation of one of the identified proteins, ubiquitin-conjugating enzyme E2 L3 (UBE2L3) is confirmed by western blotting of both nsP2-pro transfection and CHIKV natural infection, and the downregulation of UBE2L3 is additionally shown to require an enzymatically active nsP2 protease domain. Transfection of full length UBE2L3 into HEK293T/17 cells prior to CHIKV infection reduce levels of infection and E protein expression but do not alter RNA genome levels. Conclusion: These results suggest that UBE2L3 is a cellular target of the CHIKV nsP2 protease, and this possibly mediates the pathogenesis of chikungunya fever.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1002/prca.201700020