mRNA expression in mitochondria of the red alga chondrus crispus requires a unique RNA-processing mechanism, internal cleavage of upstream tRNAs at pyrimidine 48

Abstract

The hypothesis that tRNAs are involved in the maturation of the large primary transcripts of Chondrus crispus mitochondrial DNA was addressed by primer extension mapping of the transcript 5' ends of the ten genes that are preceded by tRNA genes in C. crispus mitochondrial genome. Among the 12 tRNAs that were candidates as maturation signals, eight, namely tRNAArg, tRNALys, tRNAAsp, tRNAGln, tRNATrp, tRNAIle, tRNAPhe and tRNAGly, were cleaved internally upon maturation of C. crispus mitochondrial primary transcripts, all of them at the same base, invariant pyrimidine 48. Only four tRNAs departed from this pattern: tRNALeu and tRNACys, which are not maturation signals, tRNAMet, which appears to be excised as a whole from the orf94 primary trancript and tRNAAla, which is cleaved internally at positions other than Y48. Sequence comparisons between the cleaved and the uncleaved tRNAs suggest that their core tertiary structure is involved with their recognition and cleavage. However, the precursor transcripts are also processed at the 5' and 3' ends of the tRNAs to yield tRNA molecules that are stable and functional in translation. This indicates that two different RNA processing mechanisms coexist in C. crispus mitochondria, one required for the production of functional tRNAs and the other for the processing of mRNAs.