title: Proteomic profiling identifies the SIM-associated complex of KSHV-encoded LANA.
abstract: The Kaposi's sarcoma-associated herpesvirus (KSHV) encoded latent nuclear antigen latency-associated nuclear antigen (LANA) plays an essential role in viral episome maintenance. LANA also contributes to DNA replication and tumorigenesis during latency. Recent studies suggested that LANA was involved in regulation of SUMOylation, which results in chromatin silencing. To examine the pleiotropic effects of LANA protein on host cell gene expression, we utilized MS analysis to identify cellular proteins associated with the small ubiquitin related modifier (SUMO) interacting motif of LANA (LANA(SIM)). In addition to the six bands identified as substantially associated with LANA(SIM), 151 proteins were positively identified by MS/MS analysis. Compared with previous proteomic analysis of the N- and C-truncated mutants of LANA (LANA(NC)), our results revealed that a complex of specific proteins with relatively high SUMOylation and SIM motifs is associated with LANA(SIM). Intriguingly, consistent with our previous report that identified KAP1 as a key component, the in vitro SUMO-2-modified isoform has a substantially higher affinity with LANA(SIM) than the SUMO-1-modified isoform. Moreover, via cluster and pathway analysis, we proposed a hypothetical model for the LANA(SIM) regulatory circuit involving aberrant SUMOylation of cell cycle (particular mitotic), DNA unwinding and replication, and pre-mRNA/mRNA processing related proteins. This study provides a SUMOylated and non-SUMOylated proteome profile of LANA(SIM) -associated complex and facilitates our understanding that viral-mediated gene regulation through SUMOylation is important for Kaposi's sarcoma-associated herpesvirus persistence and pathogenesis.
authors.affiliation: MOE& MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai, P. R. China.
authors.forename: Jin
authors.initials: J
authors.lastname: Gan
authors.affiliation: MOE& MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai, P. R. China.
authors.forename: Chong
authors.initials: C
authors.lastname: Wang
authors.affiliation: ShengYushou Center of Cell Biology and Immunology, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, P. R. China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Department of Gynecology, Key laboratory of AIDS immunology of Ministry of Health, First Affiliated Hospital of China Medical University, Shenyang, P. R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Biochemistry and Center for Genome Science, The University of Hong Kong, Hong Kong, P. R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.affiliation: MOE& MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai, P. R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhu
authors.affiliation: MOE& MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai, P. R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Ding
authors.affiliation: Department of Gynecology, Key laboratory of AIDS immunology of Ministry of Health, First Affiliated Hospital of China Medical University, Shenyang, P. R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Shang
authors.affiliation: Department of Biochemistry and Center for Genome Science, The University of Hong Kong, Hong Kong, P. R. China.
authors.forename: Junwen
authors.initials: J
authors.lastname: Wang
authors.affiliation: ShengYushou Center of Cell Biology and Immunology, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, P. R. China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wei
authors.affiliation: MOE& MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai, P. R. China.
authors.forename: Qiliang
authors.initials: Q
authors.lastname: Cai
authors.affiliation: Department of Microbiology and Abramson Comprehensive Cancer Center, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Erle S
authors.initials: ES
authors.lastname: Robertson
mesh_terms: D000956:Antigens, Viral / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004261:DNA Replication; D005347:Fibroblasts / Q000378:metabolism* / Q000821:virology; D006566:Herpesviridae Infections / Q000378:metabolism* / Q000821:virology; D019288:Herpesvirus 8, Human / Q000502:physiology*; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D060066:Protein Interaction Maps; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D058207:Sumoylation; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D009687:Nuclear Proteins; D020543:Proteome; D025841:Small Ubiquitin-Related Modifier Proteins; D014764:Viral Proteins; C109540:latency-associated nuclear antigen
keywords: KSHV; LANA; Microbiology; SUMO-interacting motif