title: The reversible condensation and expansion of the rotavirus genome.
abstract: Understanding the structural organization of the genome is particularly relevant in segmented double-stranded RNA viruses, which exhibit endogenous transcription activity. These viruses are molecular machines capable of repeated cycles of transcription within the intact capsid. Rotavirus, a major cause of infantile gastroenteritis, is a prototypical segmented double-stranded RNA virus. From our three-dimensional structural analyses of rotavirus examined under various chemical conditions using electron cryomicroscopy, we show here that the viral genome exhibits a remarkable conformational flexibility by reversibly changing its packaging density. In the presence of ammonium ions at high pH, the genome condenses to a radius of approximately 180 A from approximately 220 A. Upon returning to physiological conditions, the genome re-expands and fully maintains its transcriptional properties. These studies provide further insights into the genome organization and suggest that the observed isometric and concentric nature of the condensation is due to strong interactions between the genome core and the transcription enzymes anchored to the capsid inner surface. The ability of the genome to condense beyond what is normally observed in the native virus indicates that the negative charges on the RNA in the native state may be only partially neutralized. Partial neutralization may be required to maintain appropriate interstrand spacing for templates to move around the enzyme complexes during transcription. Genome condensation was not observed either with increased cation concentrations at normal pH or at high pH without ammonium ions. This finding indicates that the observed genome condensation is a synergistic effect of hydroxyl and ammonium ions involving disruption of protein-RNA interactions that perhaps facilitate further charge neutralization and consequent reduction in the interstrand spacing.
authors.affiliation: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Pesavento
authors.forename: J A
authors.initials: JA
authors.lastname: Lawton
authors.forename: M E
authors.initials: ME
authors.lastname: Estes
authors.forename: B V
authors.initials: BV
authors.lastname: Venkataram Prasad
mesh_terms: D000818:Animals; D002460:Cell Line; D020285:Cryoelectron Microscopy / Q000379:methods; D003470:Culture Media; D016679:Genome, Viral; D006863:Hydrogen-Ion Concentration; D009690:Nucleic Acid Conformation; D000644:Quaternary Ammonium Compounds; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000648:ultrastructure*; D012367:RNA, Viral / Q000737:chemistry / Q000648:ultrastructure*; D012401:Rotavirus / Q000235:genetics* / Q000648:ultrastructure; D014158:Transcription, Genetic
chemical_list: D003470:Culture Media; D000644:Quaternary Ammonium Compounds; D012330:RNA, Double-Stranded; D012367:RNA, Viral