Plant cytosolic tRNAHis possesses an exceptional C54 in the canonical TPsiC loop

A nuclear gene coding for tRNAHis from Arabidopsis has been reported to contain C54in the TPsiC loop, although the corresponding nucleotide is an invariant U or a derivative in nearly all other tRNAs. The only previously reported plant cytosolic tRNAHis sequence, from lupin, has U54. To re-examine plant cytosolic tRNAsHis and their genes we have used DNA and RNA sequence analyses, restriction enzyme digestion of PCR-amplified tRNA genes, RNA hybridization and in vivo aminoacylation assays. Our results suggest that Arabidopsis nuclear tRNAHis genes ubiquitously contain C54, as do those from tobacco, lupin and pea. The C54 nucleotide is maintained in the mature tRNAHis, which is aminoacylated in vivo , but to a relatively low level compared with other tRNAs examined. Finally, it was shown that an Arabidopsis tRNAHis gene with T54in place of C54 is over 5-fold more transcriptionally active than the wild-type gene using an in vitro system derived from plant nuclei. A possible role for this apparently sub-optimal tRNAHis sequence is suggested.