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Regulation of the Replication of Influenza Virus RNA Segments: Partial Suppression of Protein Synthesis Restores the ‘Early’ Replication Pattern

The D. I. Ivanovsky Institute of Virology of the Academy of Medical Sciences, Gamaleya Street, 16, 123098 Moscow, U.S.S.R.

The replication of influenza virus RNA was monitored by RNA-RNA hybridization with subsequent analysis of hybrid duplexes, as well as by immunosorbtion of viral nucleocapsids from extracts of [³H]uridine-labelled cells followed by the isolation and characterization of nucleocapsid-associated RNA. The nucleocapsid-associated RNA preparations contained mostly negative-strand genomic RNA. Electrophoresis of the hybrid RNA duplexes or single-stranded nucleocapsid-associated RNA in polyacrylamide gel revealed an ‘early’ replication pattern, with a predominance of the NP and NS gene segments, in cells labelled from 0 to 1 h post-infection. At later stages of infection the pattern changed to the ‘late’ one, with the M gene segment in excess of NS, and the NP gene no longer predominant. Cycloheximide added as late as 2 or 3 h post-infection suppressed RNA replication. Moderate concentrations of cycloheximide inhibited the replication of NS and NP gene segments to a lesser degree than the replication of the other RNA segments, thus restoring the ‘early’ replication pattern. Cycloheximide treatment resulted in a slight increase in the percentage of positive strands in nucleocapsid-associated RNA. The role of protein synthesis in the transition from the ‘early’ to the ‘late’ pattern of influenza virus RNA synthesis is discussed.

Keywords: influenza A virus, RNA segments, replication

Received 19 March 1987; accepted 9 July 1987.