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. 2005 Aug 2;33(13):4368–4376. doi: 10.1093/nar/gki752

Figure 4.

Figure 4

(A) Assay of RNase M5 mutant activities in the presence of ribosomal protein L18. The migration position of the precursor species (p5S), mature 5S RNA (5S), 3′ processed fragment (3′) and 5′ processed fragment (5′) are indicated to the left of the autoradiogram. Mutants were assayed at 200, 20, 2 and 0.2 ng/μl and in the presence of 0.2 ng/μl B.subtilis L18. The direction of the enzyme concentration gradient is indicated by the right-angled triangles at the top of the gel. At the highest enzyme concentration, the precursor band was shifted towards the top of the gel. (B) Assay of RNase M5 mutant activities in the presence of DMSO. Assay conditions were as in (A) except that L18 was replaced by 30% DMSO. (C) Histogram of specific activities of RNase M5 mutants relative to wild-type RNase M5. Wild-type specific activity was 5.17 pmol 5S rRNA generated per μg RNase M5 over the 15 min assay period and was set at 100%. Specific activities were calculated at the lowest dilution of the enzyme to give visible 5S rRNA processing on polyacrylamide gels and are the average of three independent assays.