title: Rubella virus P90 associates with the cytokinesis regulatory protein Citron-K kinase and the viral infection and constitutive expression of P90 protein both induce cell cycle arrest following S phase in cell culture.
abstract: In utero infection of developing fetus by Rubella virus (RV) causes cell division inhibition of critical precursor cells in organogenesis, CNS-associated birth defects and induction of apoptosis in cell culture. The underlying mechanisms of RV-induced congenital abnormalities are not known. Here, we identified a novel interaction between RV replicase P90 protein and a cytokinesis-regulatory protein, the Citron-K kinase (CK), in a yeast two-hybrid cDNA library screen. Aberrations in cytokinesis and subsequent apoptosis do occur in specific cell types when the CK gene is knocked out or, its regulatory function is perturbed. Our analysis found that full-length P90 binds CK and in RV-infected cells P90 colocalizes with CK in the cytoplasm. Furthermore, during RV infection as well as cellular expression of P90 alone, we identified a discrete subpopulation of cells containing 4N DNA content, indicating that these cells are arrested in the cell cycle following S phase, suggesting that cellular expression of viral P90 during RV infection perturbs cytokinesis. Previous reports by others established that RV infection leads to apoptosis in cell culture. These observations together taken to the fetal organogenesis level, favor the idea that RV P90, by binding to cellular CK, invokes cell cycle aberrations resulting in the cell- and organ-specific growth inhibition and programmed cell death during RV infection in utero, which commonly is referred to as RV-induced teratogenesis.
authors.affiliation: Section of Viral Pathogenesis and Adverse Reactions, Laboratory of Pediatric and Respiratory Viral Diseases, Center for Biologics Evaluation and Research, Bethesda, Maryland 20892, USA. Atreya@cber.fda.gov
authors.forename: C D
authors.initials: CD
authors.lastname: Atreya
authors.forename: S
authors.initials: S
authors.lastname: Kulkarni
authors.forename: K V K
authors.initials: KV
authors.lastname: Mohan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D012409:Rubella / Q000151:congenital / Q000821:virology; D012412:Rubella virus / Q000737:chemistry / Q000378:metabolism*; D016196:S Phase; D016415:Sequence Alignment; D020798:Two-Hybrid System Techniques; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000378:metabolism*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D017361:Viral Nonstructural Proteins; C115635:citron-kinase; D017346:Protein Serine-Threonine Kinases