title: An intein-mediated modulation of protein stability system and its application to study human cytomegalovirus essential gene function.
abstract: Functional analysis of the essential proteins encoded by human cytomegalovirus (HCMV) is hindered by the lack of complementing systems. To overcome this difficulty, we have established a novel approach, termed the intein-mediated modulation of protein stability (imPS), in which a destabilizing domain and part of a split intein are fused to the essential protein. The growth of the mutant virus can then be regulated by the degradation and splicing of the protein. We found that an ultrafast gp41-1 split intein was able to rescue or degrade the protein of interest (POI) by removing or adding a strong degron through protein splicing. As a result, the function of the POI was turned on or off during the process. Using HCMV essential gene IE1/IE2, we confirmed that imPS worked remarkably well in conditionally regulating protein stability during viral infection. This conditional approach is likely to be applicable for dissecting the gene functions of HCMV or other viruses.
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.
authors.forename: Deng
authors.initials: D
authors.lastname: Pan
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.
authors.forename: Baoqin
authors.initials: B
authors.lastname: Xuan
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.
authors.forename: Yamei
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.|Institutes for Advanced Interdisciplinary Research, East China Normal University, Shanghai, China.
authors.forename: Shaowu
authors.initials: S
authors.lastname: Huang
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.|Jiangsu Key laboratory of infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou, China.
authors.forename: Maorong
authors.initials: M
authors.lastname: Xie
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.|Jiangsu Key laboratory of infection and Immunity, Institutes of Biology and Medical Sciences, Soochow University, Suzhou, China.
authors.forename: Yadan
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.
authors.forename: Wenjia
authors.initials: W
authors.lastname: Xu
authors.affiliation: Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology &Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Shanghai, China.
authors.forename: Zhikang
authors.initials: Z
authors.lastname: Qian
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics* / Q000254:growth & development; D020043:Genes, Essential; D005814:Genes, Viral; D057809:HEK293 Cells; D006801:Humans; D047668:Inteins / Q000235:genetics; D050296:Microbial Viability; D008967:Molecular Biology / Q000379:methods*; D055550:Protein Stability; D014773:Virology / Q000379:methods*