Inhibition of dengue virus translation and RNA synthesis by a morpholino oligomer targeted to the top of the terminal 3' stem-loop structure.
Dengue virus (DEN) is a major public health problem worldwide and causes a spectrum of diseases, for which no antiviral treatments exist. Peptide-conjugated phosphorodiamidate morpholino oligomers (P-PMOs) complementary to the DEN 5' stem-loop (5'SL) and to the DEN 3' cyclization sequence (3'CS) inhibit DEN replication, presumably by blocking critical RNA-RNA or RNA-protein interactions involved in viral translation and/or RNA synthesis. Here, a third P-PMO, complementary to the top of the 3' stem-loop (3'SLT), inhibited DEN replication in BHK cells. Using a novel DEN2 reporter replicon and a DEN2 reporter mRNA, we determined that the 5'SL P-PMO inhibited viral translation, the 3'CS P-PMO blocked viral RNA synthesis but not viral translation, and the 3'SLT P-PMO inhibited both viral translation and RNA synthesis. These results show that the 3'CS and the 3'SL domains regulate DEN translation and RNA synthesis and further demonstrate that P-PMOs are potentially useful as antiviral agents.
Holden
Katherine Lynn
KL
Division of Infectious Diseases, School of Public Health, University of California at Berkeley, 140 Warren Hall, Berkeley, CA 94720-7360, USA.
Stein
David A
DA
Pierson
Theodore C
TC
Ahmed
Asim A
AA
Clyde
Karen
K
Iversen
Patrick L
PL
Harris
Eva
E
D020413:3' Untranslated Regions / Q000737:chemistry / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003716:Dengue Virus / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D017930:Genes, Reporter; D009690:Nucleic Acid Conformation; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects*; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry*; D013379:Substrate Specificity; D014779:Virus Replication / Q000187:drug effects
D020413:3' Untranslated Regions; D009842:Oligopeptides; D012367:RNA, Viral