Effects of (hydroxymethyl)trimethylpsoralen on structure and function of bacteriophage MS2 ribonucleic acid

Treatment of bacteriophage MS2 with 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen and 360-nm light caused a dose-dependent decline in the infectivity of the virus. Covalent photobinding of a single psoralen molecule on the phage genome was a lethal event. Ribonucleic acid (RNA) extracted from psoralen and light-treated virus had a dose-dependent 385-nm fluorescence emission but was unaltered in its physical properties compared to control RNA samples. Phage adsorption and penetration in Escherichia coli host cells were unaffected, but in vivo replication of the treated virus was affected to the same extent as infectivity. The cell-free translational activity of the MS2 RNA was also severely reduced after psoralen and light treatment of the phage. Examination of the in vitro translation products revealed that the synthesis of the viral replicase protein was most substantially affected. Psoralen treatment of purified, protein-free MS2 RNA promoted an even greater reduction in cell-free synthesis of all viral proteins. This difference in translational function was consistent with the observation that virion-free RNA bound approximately 4 times as much psoralen as did RNA treated within the phage capsid. It was concluded that the replicase gene is the most sensitive region of the viral RNA molecule for psoralen binding.