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. 2002 Jul;22(14):5248–5256. doi: 10.1128/MCB.22.14.5248-5256.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 2. — Processing of tRNAs is affected in lsm mutants. (A to F) Analysis of tRNA processing in GAL::lsm and lsm-Δ strains. (G and H) Analysis of processing of tRNA^Tyr and in GAL::lsm3 strain. (I to K) Analysis of tRNA processing in lsm2^ts and lsm5^ts strains. Strains carrying GAL-regulated HA-tagged constructs (GAL::lsm; A to F, lanes 3 to 8, and G and H, lanes 3 to 6) and the BMA64 wild-type strain (A to F, lanes 1 and 2, and G and H, lanes 1 and 2) were grown in permissive RSG medium (0 h) and transferred to repressive glucose medium at 30°C for the times indicated. Strains with deletions of Lsm6p or Lsm1p (A to F, lanes 11 to 16), the temperature-sensitive lsm2^ts and lsm5^ts strains (I to K, lanes 3 to 8), and the wild-type strain (A to F, lanes 9 and 10, and I to K, lanes 1 and 2) were pregrown at 23°C (0 h) and transferred to 37°C for the times indicated. RNA was separated on a 6% polyacrylamide gel and hybridized with oligonucleotide probes. Names of tRNAs are on the left; schematic representations of tRNA precursors, intermediates, and mature species are shown between the two columns (A to F) or on the right (G to K); and probe names are in parentheses. RNA precursors marked with asterisks represent species predicted to be 3′ extended beyond the wild-type transcripts. tRNA^Pro precursors designated by a and b represent two forms of PT (32). The minor size heterogeneity observed for pre-tRNAs and the mature tRNAs may result from the presence or absence of the 3′-end -CCA_OH tail. WT, wild type.

Inline graphic — Processing of tRNAs is affected in lsm mutants. (A to F) Analysis of tRNA processing in GAL::lsm and lsm-Δ strains. (G and H) Analysis of processing of tRNA^Tyr and in GAL::lsm3 strain. (I to K) Analysis of tRNA processing in lsm2^ts and lsm5^ts strains. Strains carrying GAL-regulated HA-tagged constructs (GAL::lsm; A to F, lanes 3 to 8, and G and H, lanes 3 to 6) and the BMA64 wild-type strain (A to F, lanes 1 and 2, and G and H, lanes 1 and 2) were grown in permissive RSG medium (0 h) and transferred to repressive glucose medium at 30°C for the times indicated. Strains with deletions of Lsm6p or Lsm1p (A to F, lanes 11 to 16), the temperature-sensitive lsm2^ts and lsm5^ts strains (I to K, lanes 3 to 8), and the wild-type strain (A to F, lanes 9 and 10, and I to K, lanes 1 and 2) were pregrown at 23°C (0 h) and transferred to 37°C for the times indicated. RNA was separated on a 6% polyacrylamide gel and hybridized with oligonucleotide probes. Names of tRNAs are on the left; schematic representations of tRNA precursors, intermediates, and mature species are shown between the two columns (A to F) or on the right (G to K); and probe names are in parentheses. RNA precursors marked with asterisks represent species predicted to be 3′ extended beyond the wild-type transcripts. tRNA^Pro precursors designated by a and b represent two forms of PT (32). The minor size heterogeneity observed for pre-tRNAs and the mature tRNAs may result from the presence or absence of the 3′-end -CCA_OH tail. WT, wild type.