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. 2007 Nov 2;7(1):86–101. doi: 10.1128/EC.00215-07

FIG. 7.

FIG. 7.

Comparison of the levels of Nm modifications in bloodstream-stage (BS) versus procyclic-stage (PS) trypanosomes. Total RNA (8 μg) was prepared from T. brucei procyclic and bloodstream forms and was subjected to primer extension with radiolabeled oligonucleotides, performed as described for Fig. 2. Partial sequences are given, and the modified nucleotides are marked by solid arrows and circles. To control for the level of RNA in each sample, primer extension was performed with a primer specific to U4 snRNA. (a) Mapping of Nms on LSU rRNA (3′ half) using primer 2 LSU-AS 1098-1117. The modified nucleotides that are developmentally regulated are circled. (b) The level of TB5Cs1C1 snoRNA, which directs the Am1031 modification, was analyzed by primer extension using a radiolabeled oligonucleotide against TB5Cs1C1 snoRNA. U6 snRNA served as a control for the level of RNA in each sample. (c) Positions of the modified nucleotides on the secondary structure of T. brucei LSU 3′ rRNA. Nm modifications in T. brucei (T), S. cerevisiae (Y), the plant A. thaliana (P), and humans (H) are indicated.