Nipah virus conforms to the rule of six in a minigenome replication assay.
To study the replication of Nipah virus (NiV), a minigenome replication assay that does not require the use of infectious virus was developed. The minigenome was constructed to encode a NiV vRNA analogue containing the gene for chloramphenicol acetyltransferase (CAT) under the control of putative NiV transcription motifs and flanked by the NiV genomic termini. CAT protein was detected only when plasmids encoding the NiV minigenome, nucleocapsid protein (N), phosphoprotein (P) and polymerase protein (L) were transfected into CV1 cells. To determine whether NiV conforms to the rule of six, a series of plasmids encoding minigenomes that differed in length by a single nucleotide was tested in the replication assay. CAT production was detected only with the minigenome whose length was an even multiple of six. The replication assay was also used to show that the N, P and L proteins of NiV recognize cis-acting sequences in the genomic termini of Hendra virus (HeV) but not measles virus. While these results suggest that NiV uses a replication strategy that is similar to those of other paramyxoviruses, they also support the inclusion of NiV and HeV in a separate genus within the subfamily Paramyxovirinae.
Halpin
Kim
K
Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-C22, Atlanta, GA 30333, USA.
Bankamp
Bettina
B
Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-C22, Atlanta, GA 30333, USA.
Harcourt
Brian H
BH
Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-C22, Atlanta, GA 30333, USA.
Bellini
William J
WJ
Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-C22, Atlanta, GA 30333, USA.
Rota
Paul A
PA
Measles Virus Section, National Center for Infectious Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, MS-C22, Atlanta, GA 30333, USA.
D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D017930:Genes, Reporter; D016679:Genome, Viral; D006801:Humans; D045405:Nipah Virus / Q000235:genetics* / Q000502:physiology; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D013552:Swine; D013553:Swine Diseases / Q000821:virology; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication / Q000235:genetics; D015047:Zoonoses
D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins