Open reading frame in rotavirus mRNA specifically promotes synthesis of double-stranded RNA: Template size also affects replication efficiency

Citation
Jt. Patton et al., Open reading frame in rotavirus mRNA specifically promotes synthesis of double-stranded RNA: Template size also affects replication efficiency, VIROLOGY, 264(1), 1999, pp. 167-180
Citations number
34
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Microbiology
Journal title
VIROLOGY
ISSN journal
0042-6822 → ACNP
Volume
264
Issue
1
Year of publication
1999
Pages
167 - 180
Database
ISI
SICI code
0042-6822(19991110)264:1<167:ORFIRM>2.0.ZU;2-C
Abstract
The 11 rotavirus mRNAs are capped, but not polyadenylated, have a high AU c ontent, and serve as templates for the synthesis of double-stranded (ds)RNA . Earlier studies using a cell-free replication system showed that the 5'- and 3'-untranslated regions (UTRs) of the mRNAs have cis-acting signals tha t promote minus-strand synthesis. To identify additional factors that affec t RNA replication, chimeric RNAs were made that consisted of portions of th e gene 8 mRNA of SA11 rotavirus and of the gene for green fluorescent prote in (gfp) or for the N protein of respiratory syncytial virus. Analysis of t he chimeras in the cell-free replication system under noncompetitive condit ions showed that the open reading frame (ORF) of viral mRNAs contains infor mation that specifically promotes minus-strand synthesis. Results were also obtained indicating that a high AU content may increase the replication ef ficiency of RNAs and that, in general, an inverse correlation exists betwee n replication efficiency and the length of the RNA template. Replication as says performed under competitive conditions showed that nonviral RNAs can i nterfere significantly with the replication of viral mRNAs, mostly likely b y sequestering nonspecific RNA-binding proteins that are of limited concent ration in the replication system and that are essential for dsRNA synthesis . In summary, rotavirus dsRNA synthesis is affected by many factors includi ng cis-acting replication signals located in the 5'-UTR, 3'-UTR, and ORF of the mRNA as well as the size and possibly the AU content of the mRNA. (C) 1999 Academic Press.