Effect of RNase III on the size of bacteriophage T7 lysozyme mRNA

F S Hagen, E T Young
1978 Journal of Virology  
The size of lysozyme mRNA from T7-infected E. coli RNase III' and RNase III-strains was analyzed by sucrose gradient sedimentation, dimethylsulfoxide (Me2SO) sucrose gradient sedimentation, and preparative gel electrophoresis. Each technique revealed a similar size distribution of multiple lysozyme mRNA's. Analysis by preparative gel electrophoresis of RNA extracted after infection of Escherichia coli B8t (RNase III') separated lysozyme mRNA into six peaks of activity ranging in size from 0.2 x
more » ... 106 to 1.9 x 106 daltons. Four well-resolved major peaks of activity were detected, having apparent molecular weights of approximately 0.61 x 106, 0.76 x 10", 0.92 x 10", and 1.3 x 106. A broad band of activity, with a molecular weight range from 0.2 x 106 to 0.37 x 106, was also present, and a sixth peak of activity was sometimes observed that migrates with a mobility corresponding to a molecular weight of 1.9 x 106. Judging from their molecular weight as estimated by electrophoresis, most, if not all, of the lysozyme mRNA's were polycistronic. The RNA extracted after infection of an RNase IIIhost contained a more heterogeneous collection of lysozyme mRNA's. In addition to lysozyme mRNA activity on RNAs with molecular weights between 0.2 x 106 and 1.9 x 106, RNA species with molecular weights estimated at 4 x 10' to 5 x 106 were also detected. The data indicate that RNase III processes at least some of the primary lysozyme transcripts. The multiple lysozyme mRNA's represent discrete RNA species rather than aggregates because analysis of the size of lysozyme mRNA under completely denaturing conditions, in Me2SO, produced a similar size distribution of lysozyme mRNAs. Also, treatment of RNA with 90% Me2SO, which separates the strands of a completely double-stranded RNA, did not significantly alter the electrophoretic mobility of the lysozyme mRNA.
doi:10.1128/jvi.26.3.783-792.1978 fatcat:rvwe4awcm5btnjwe3j3elvi7ha