Inhibition of replication of human respiratory syncytial virus by 6-diazo-5-oxo-L-norleucine.
The effect of 6-diazo-5-oxo-L-norleucine (L-DON), a glutamine analog, on RSV replication was studied. At a concentration of 0.01 mM L-DON, 99% of RSV replication in treated CV-1 cells was inhibited. At this concentration of L-DON, the level of cellular protein synthesis was identical to untreated control cells. Trypan blue staining revealed that all the cells remained viable even at concentrations of L-DON as high as 10 mM. In addition, L-DON added as late as 24 h post infection can effectively suppress viral replication. Analysis of viral mRNA levels by Northern blot revealed that secondary transcription and subsequent steps in the virus life cycle were inhibited. Immunoprecipitation of viral proteins from drug treated or untreated cultures showed that synthesis of all viral proteins was drastically reduced by L-DON, with a slightly greater inhibition of viral glycoproteins. Furthermore, immunofluorescent staining showed that drug treated cells expressed both F and N proteins and that F was inserted into the membrane as the native F protein.
Huang
R C
RC
Institute of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106, USA.
Panin
M
M
Romito
R R
RR
Huang
Y T
YT
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D003980:Diazooxonorleucine / Q000494:pharmacology*; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D018113:Respiratory Syncytial Virus, Human / Q000187:drug effects* / Q000502:physiology; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D003980:Diazooxonorleucine