The activity of Sendai virus genomic and antigenomic promoters requires a second element past the leader template regions: a motif (GNNNNN)3 is essential for replication

Article Article scientifique Article The activity of Sendai virus genomic and antigenomic promoters requires a second element past the leader template regions: a motif (GNNNNN)3 is essential for replication eng 1998 author Tapparel Caroline 0000-0002-0411-6567 27039 author Maurice Diane Barthelemie Emilie 28268 author Roux Laurent 24751 3117-3128 10.1128/JVI.72.4.3117-3128.1998 9525637 0022-538X The paramyxovirus genome, a nonsegmented, negative-polarity, single-stranded RNA of approximately 15 kb, contains six transcription units flanked at the 3' and 5' ends by a short (approximately 50- to 60-nucleotide) extracistronic sequence, dubbed the positive and negative leader regions. These leader template regions, present at the 3' end of the genome and the antigenome, have been shown to contain essential signals governing RNA replication activity. Whether they are sufficient to promote replication is still open to question. By using a series of Sendai virus defective interfering RNAs carrying a nested set of deletions in the promoter regions, it is shown here that for both the genomic and antigenomic promoters, a 3'-end RNA sequence of 96 nucleotides is required to allow replication. Sequence comparison of active and inactive promoters led to the identification of a set of three nucleotide hexamers (nucleotides 79 to 84, 85 to 90, and 91 to 96) containing a repeated motif RXXYXX [shown as 5'-3' positive-strand]. Sequential mutation of each hexamer into its complementary sequence confirmed their essential role. The three hexamers are required, and their relative positioning is important, since displacing them by 6 nucleotides destroyed promoter function. RNAs carrying degenerate nucleotides in the three hexamers were used as replication templates. They led to the selection of actively replicating RNA species exclusively carrying the basic motif (GNNNNN)3 from nucleotides 79 to 96. These results clearly show that, apart from the region from nucleotides 1 to 31, previously identified as governing Sendai virus replication activity, a second element, spanning at the most nucleotides 79 to 96, appears essential. Thus, the paramyxovirus replication promoters are not confined to the leader template regions, as seems to be the case for the rhabdoviruses. Binding Sites Genome Viral HeLa Cells Humans Promoter Regions Genetic RNA Messenger RNA Viral/biosynthesis Respirovirus/genetics Templates Genetic Tumor Cells Cultured Dewey 616 54 Département de microbiologie et médecine moléculaire 21B false 931 Identification des facteurs clés impliqués dans la pathogénèse des rhinovirus et entérovirus 953 Journal of Virology 72 4 82ec8d4c-484e-47a8-ab80-0b0ab3e6ccaf Article (Published version) Public jv003117.pdf 2052487 application/pdf OA OTHER openAccess Faculté de médecine / Section de médecine fondamentale / Département de microbiologie et médecine moléculaire

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The activity of Sendai virus genomic and antigenomic promoters requires a second element past the leader template regions: a motif (GNNNNN)3 is essential for replication	Journal of Virology			1998	610	228

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Maurice, Diane Barthelemie Emilie