title: Transmissible gastroenteritis coronavirus genome packaging signal is located at the 5' end of the genome and promotes viral RNA incorporation into virions in a replication-independent process.
abstract: Preferential RNA packaging in coronaviruses involves the recognition of viral genomic RNA, a crucial process for viral particle morphogenesis mediated by RNA-specific sequences, known as packaging signals. An essential packaging signal component of transmissible gastroenteritis coronavirus (TGEV) has been further delimited to the first 598 nucleotides (nt) from the 5' end of its RNA genome, by using recombinant viruses transcribing subgenomic mRNA that included potential packaging signals. The integrity of the entire sequence domain was necessary because deletion of any of the five structural motifs defined within this region abrogated specific packaging of this viral RNA. One of these RNA motifs was the stem-loop SL5, a highly conserved motif in coronaviruses located at nucleotide positions 106 to 136. Partial deletion or point mutations within this motif also abrogated packaging. Using TGEV-derived defective minigenomes replicated in trans by a helper virus, we have shown that TGEV RNA packaging is a replication-independent process. Furthermore, the last 494 nt of the genomic 3' end were not essential for packaging, although this region increased packaging efficiency. TGEV RNA sequences identified as necessary for viral genome packaging were not sufficient to direct packaging of a heterologous sequence derived from the green fluorescent protein gene. These results indicated that TGEV genome packaging is a complex process involving many factors in addition to the identified RNA packaging signal. The identification of well-defined RNA motifs within the TGEV RNA genome that are essential for packaging will be useful for designing packaging-deficient biosafe coronavirus-derived vectors and providing new targets for antiviral therapies.
authors: Department of Molecular and Cell Biology, National Center of Biotechnology (CNB-CSIC), Campus de la Universidad Autonoma de Madrid, Madrid, Spain.
authors: Lucia
authors: L
authors: Morales
authors: Pedro A
authors: PA
authors: Mateos-Gomez
authors: Carmen
authors: C
authors: Capiscol
authors: Lorena
authors: L
authors: del Palacio
authors: Luis
authors: L
authors: Enjuanes
authors: Isabel
authors: I
authors: Sola
mesh_terms: D000818:Animals; D002460:Cell Line; D017934:Coronavirus / Q000235:genetics / Q000502:physiology*; D004252:DNA Mutational Analysis; D017354:Point Mutation; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D014771:Virion / Q000378:metabolism; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D012367:RNA, Viral
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