title: Glutathionylation of chikungunya nsP2 protein affects protease activity.
abstract: Chikungunya fever is an emerging disease caused by the chikungunya virus and is now being spread worldwide by the mosquito Aedes albopictus. The infection can cause a persistent severe joint pain and recent reports link high levels of viremia to neuropathologies and fatalities. The viral protein nsP2 is a multifunctional enzyme that plays several critical roles in virus replication. Virus infection induces oxidative stress in host cells which the virus utilizes to aid viral propagation. Cellular oxidative stress also triggers glutathionylation which is a post-translational protein modification that can modulate physiological roles of affected proteins.
  
  
  
  The nsP2 protease is necessary for processing of the virus nonstructural polyprotein generated during replication. We use the recombinant nsP2 protein to measure protease activity before and after glutathionylation. Mass spectrometry allowed the identification of the glutathione-modified cysteines. Using immunoblots, we show that the glutathionylation of nsP2 occurs in virus-infected cells.
  
  
  
  We show that in virus-infected cells, the chikungunya nsP2 can be glutathionylated and we show this modification can impact on the protease activity. We also identify 6 cysteine residues that are glutathionylated of the 20 cysteines in the protein.
  
  
  
  The virus-induced oxidative stress causes modification of viral proteins which appears to modulate virus protein function.
  
  
  
  Viruses generate oxidative stress to regulate and hijack host cell systems and this environment also appears to modulate virus protein function. This may be a general target for intervention in viral pathogenesis.
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Thailand.
authors.forename: Chonticha
authors.initials: C
authors.lastname: Saisawang
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Thailand.
authors.forename: Atichat
authors.initials: A
authors.lastname: Kuadkitkan
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Thailand; Center for Emerging and Neglected Infectious Diseases, Mahidol University, Thailand.
authors.forename: Duncan R
authors.initials: DR
authors.lastname: Smith
authors.affiliation: Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand; Center for Emerging and Neglected Infectious Diseases, Mahidol University, Thailand.
authors.forename: Sukathida
authors.initials: S
authors.lastname: Ubol
authors.affiliation: Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Thailand. Electronic address: albertketterman@yahoo.com.
authors.forename: Albert J
authors.initials: AJ
authors.lastname: Ketterman
mesh_terms: D002460:Cell Line; D065632:Chikungunya Fever / Q000378:metabolism / Q000821:virology; D002646:Chikungunya virus / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018384:Oxidative Stress / Q000502:physiology; D010447:Peptide Hydrolases / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology
chemical_list: D017361:Viral Nonstructural Proteins; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D005978:Glutathione; D003545:Cysteine
keywords: Alphavirus; Chikungunya virus; Enzyme modulation; Glutathionylation; Nonstructural protein 2 (nsP2)