NS1 interaction with CKII alpha: novel protein complex mediating parvovirus-induced cytotoxicity.
During a productive infection, the prototype strain of the parvovirus minute virus of mice (MVMp) induces dramatic morphological alterations in permissive A9 fibroblasts, culminating in cell lysis at the end of infection. These cytopathic effects (CPE) result from rearrangements and destruction of the cytoskeletal micro- and intermediate filaments, while other structures such as the nuclear lamina and particularly the microtubule network remain protected throughout the infection (J. P. F. Nüesch et al., Virology 331:159-174, 2005). In order to unravel the mechanism(s) by which parvoviruses trigger CPE, we searched for NS1 interaction partners by differential affinity chromatography, using distinct NS1 mutants debilitated specifically for this function. Thereby, we isolated an NS1 partner polypeptide, whose interaction with NS1 correlated with the competence of the viral product for CPE induction, and further identified it by tandem mass spectrometry and Western blotting analyses to consist of the catalytic subunit of casein kinase II, CKIIalpha. This interaction of NS1 with CKIIalpha suggested interference by the viral protein with intracellular signaling. Using permanent cell lines expressing dominant-negative CKIIalpha mutants, we were able to show that this kinase activity was indeed specifically involved in parvoviral CPE and progeny particle release. Furthermore, the NS1/CKIIalpha complex proved to be able to specifically phosphorylate viral capsids, indicating a mediator function of NS1 for CKII activity and specificity, at least in vitro. Altogether our data suggest that parvovirus-induced CPE is mediated by NS1 interference with intracellular CKII signaling.
Nüesch
Jürg P F
JP
Program Infection and Cancer, Abt. F010 and INSERM U701, Deutsches Krebsforschungszentrum, Heidelberg, Germany. jpf.nuesch@dkfz-heidelberg.de
Rommelaere
Jean
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D036022:Capsid Proteins / Q000378:metabolism; D047390:Casein Kinase II / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D003588:Cytopathogenic Effect, Viral / Q000276:immunology; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007424:Intracellular Fluid / Q000276:immunology / Q000378:metabolism / Q000821:virology; D051379:Mice; D008915:Minute Virus of Mice / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D015398:Signal Transduction / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000378:metabolism* / Q000502:physiology
D036022:Capsid Proteins; C106202:NS1 protein, parvovirus; D017361:Viral Nonstructural Proteins; D047390:Casein Kinase II