Characterization of the RNA chaperone activity of hantavirus nucleocapsid protein.
Hantaviruses are tripartite negative-sense RNA viruses and members of the Bunyaviridae family. The nucleocapsid (N) protein, encoded by the smallest of the three genome segments (S), has nonspecific RNA chaperone activity. This activity results in transient dissociation of misfolded RNA structures, may be required for facilitating correct higher-order RNA structure, and may function in viral genome replication. We carried out a series of experiments to further characterize the ability of N to dissociate RNA duplexes. As might be expected, N dissociated RNA duplexes but not DNA duplexes or RNA-DNA heteroduplexes. The RNA-destabilizing activity of N is ATP independent, has a pH optimum of 7.5, and has an Mg(2+) concentration optimum of 1 to 2 mM. N protein is unable to unwind the RNA duplexes that are completely double stranded. However, in the presence of an adjoining single-stranded region, helix unwinding takes place in the 3'-to-5' direction through an unknown mechanism. The N protein trimer specifically recognizes and unwinds the terminal panhandle structure in the viral RNA and remains associated with unwound 5' terminus. We suggest that hantaviral nucleocapsid protein has an active role in hantaviral replication by working cooperatively with viral RNA polymerase. After specific recognition of the panhandle structure by N protein, the unwound 5' terminus likely remains transiently bound to N protein, creating an opportunity for the viral polymerase to initiate transcription at the accessible 3' terminus.
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Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
Panganiban
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D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D018833:Chaperonins / Q000737:chemistry / Q000378:metabolism*; D016679:Genome, Viral / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000737:chemistry; D009690:Nucleic Acid Conformation; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000378:metabolism; D029261:Sin Nombre virus / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
D019590:Nucleocapsid Proteins; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D000255:Adenosine Triphosphate; D012324:RNA-Dependent RNA Polymerase; D018833:Chaperonins; D008274:Magnesium