title: DNA topoisomerase 1 facilitates the transcription and replication of the Ebola virus genome.
abstract: Ebola virus (EBOV) protein L (EBOL) acts as a viral RNA-dependent RNA polymerase. To better understand the mechanisms underlying the transcription and replication of the EBOV genome, we sought to identify cellular factors involved in these processes via their coimmunoprecipitation with EBOL and by mass spectrometry. Of 65 candidate proteins identified, we focused on DNA topoisomerase 1 (TOP1), which localizes to the nucleus and unwinds helical DNA. We found that in the presence of EBOL, TOP1 colocalizes and interacts with EBOL in the cytoplasm, where transcription and replication of the EBOV genome occur. Knockdown of TOP1 markedly reduced virus replication and viral polymerase activity. We also found that the phosphodiester bridge-cleaving and recombination activities of TOP1 are required for the polymerase activity of EBOL. These results demonstrate that TOP1 is an important cellular factor for the transcription and replication of the EBOV genome and, as such, plays a key role in the EBOV life cycle.
authors: Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors: Kei
authors: K
authors: Takahashi
authors: Peter
authors: P
authors: Halfmann
authors: Masaaki
authors: M
authors: Oyama
authors: Hiroko
authors: H
authors: Kozuka-Hata
authors: Takeshi
authors: T
authors: Noda
authors: Yoshihiro
authors: Y
authors: Kawaoka
mesh_terms: D000818:Animals; D002460:Cell Line; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D029043:Ebolavirus / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D025941:Protein Interaction Mapping; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase; D004264:DNA Topoisomerases, Type I; C528909:TOP1 protein, human
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