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. 1998 Mar 17;95(6):2838–2843. doi: 10.1073/pnas.95.6.2838

Figure 3.

Figure 3

In vivo import of tRK2cuu. (a) Sequences of the tRK2 (GenBank accession no. U40828; chromosome XVI) and tRK1 (20) genes; arrows indicate sequences corresponding to oligonucleotides used for the subcloning of the two genes; 5′ and 3′ flanking regions are in italic type; underlined lowercase type shows intron sequences of the tRK2 gene; boldface type shows the “coding” sequence; anticodons are boxed. (b) Predicted secondary structure of the intron containing part of the tRK2 gene; intron sequences are outlined with plain arrows indicating sites of cleavage; the shadowed arrow indicates the U34 to C replacement in tRK2CUU; the anticodon is boxed. (c) Northern blot hybridization of total (Cyt) and mitochondrial (Mit) tRNAs from wild-type and pTRK2-CUU transformed cells, using oligonucleotide probes specific for tRK2CUU (anti-CUU) and tRK2UUU (anti-UUU). Temperatures of washes are indicated above the autoradiographs. At 40°C, the anti-CUU probe recognizes both mutant and wild-type tRK2, whereas at 45°C, only tRK2CUU is detected. Arrows indicate the tRK2CUU (CUU) and tRK2mnmLys5s2UUU (U*UU) specific bands; 5 μg of total and 15 μg of mitochondrial tRNA were loaded in each slot. (d) Dot hybridization of total (Cyt) and mitochondrial (Mit) tRNA with tRK1, tRK2, tRK3, tRK1UUU-7, and tRK2CUU-specific probes; the amount of dotted RNA is indicated above the autoradiographs.