BMV RNA2 translation is strongly inhibited by mutations in LSM1 affecting RNA-binding or intersubunit contacts. lsm1Δ yeast strain was transformed with a plasmid transcribing BMV RNA2 together with an empty plasmid or a plasmid expressing LSM1 or the different lsm1 mutant alleles. (Upper panels) Northern blot (NB) analyses of RNA2 accumulation. Detection of the 18S ribosomal RNA was used to assure equal loading and sample quality. (Lower panels) Western blot analyses (WB) of 2a and Lsm1 expression. As a control for equal loading of total protein, expression of the phosphoglycerate kinase protein (PGK) was also analyzed. Histograms show the average and standard error of the mean of the relative accumulation of 2a protein relative to the amount of RNA2 for at least three independent colonies. The average value obtained in cells expressing LSM1 gene was set to 100.
