Characterization of N protein self-association in coronavirus ribonucleoprotein complexes.
Mouse hepatitis virus (MHV) nucleocapsid (N) protein binds to the large, single-stranded, positive-sense viral genomic RNA to form a helical nucleocapsid structure in mature virions. In addition N protein binds the intracellular form of the genomic RNA, all of the MHV subgenomic mRNAs, and expressed non-MHV RNA transcripts to form ribonucleoprotein (RNP) complexes in infected cells. Among the intracellular viral RNP complexes, only the genomic RNP complex is packaged into virus particles. The present study demonstrated that N protein in the MHV virion nucleocapsid and in the intracellular genome-length RNP complex that bound to viral envelope M protein was tightly self-associated such that its association was retained even after extensive RNase A-treatment of the RNP complexes. The RNase A-resistant tight N protein association in the virion nucleocapsid was not mediated by an intermolecular disulfide bridge between N proteins. In contrast, N protein association in the majority of the intracellular RNP complexes was susceptible to RNase A-treatment. Because the RNP complexes that specifically interact with the M protein are selectively packaged into MHV particles, the present data suggested that there was a distinct difference between N protein association in viral genomic RNP complexes that undergo packaging into virus particles and the subgenomic RNP complexes that are not packaged into MHV particles.
Narayanan
Krishna
K
Department of Microbiology and Immunology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555-1019, USA.
Kim
Kyongmin Hwang
KH
Makino
Shinji
S
D000818:Animals; D002460:Cell Line; D000086462:Coronavirus Nucleocapsid Proteins; D051379:Mice; D006517:Murine hepatitis virus / Q000378:metabolism* / Q000502:physiology; D019590:Nucleocapsid Proteins / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000502:physiology*; D012261:Ribonucleoproteins / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000378:metabolism*; D019065:Virus Assembly
D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014763:Viral Matrix Proteins