Summary
We have recently identified a tRNA gene cluster in theArabidopsis nuclear genome. One tRNASer (AGA) gene and two tRNATyr (GTA) genes occur in tandem arrangement on a 1.5 kb unit that is amplified about 20-fold at a single chromosomal site. Here we have studied the in vitro expression of seven individually cloned tRNASer genes (pAtS1 to pAtS7) derived from this cluster. Five out of the seven tRNASer genes contain point mutations in the coding region which have in part adverse effects on the expression of these genes in different cell-free systems: (i) C10 and A62 in plant tRNASer genes, which correspond to G10 and C62, respectively, in all known vertebrate tRNA genes, result in a reduced transcription efficiency in HeLa but not in yeast extract. This indicates that yeast RNA polymerase III tolerates nucleotide substitutions at positions 10 [5′ internal control region (ICR)] and 62 (3′ ICR), whereas the vertebrate RNA polymerase III requires a more stringent consensus sequence. (ii) Processing of a pre-tRNASer with a mismatch in the aminoacyl stem is impaired in HeLa, yeast and wheat germ extracts, however, a mismatch in the anticodon stem is deleterious for HeLa and wheat germ but not for yeast processing enzymes. The unexpectedly high number of potential tRNASer pseudogenes in the cluster — quite in contrast to the tRNATyr genes which mainly code for functional tRNAs — suggested that tRNASer (AGA) genes also occur elsewhere in the genome. We present evidence that single copies of tRNASer (AGA) genes do indeed exist outside the tRNA gene cluster.
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Beier, D., Beier, H. Expression of variant nuclearArabidopsis tRNASer genes and pre-tRNA maturation differ in HeLa, yeast and wheat germ extracts. Molec. Gen. Genet. 233, 201–208 (1992). https://doi.org/10.1007/BF00587580
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DOI: https://doi.org/10.1007/BF00587580