The steady-state level of mRNA from the Ogura cytoplasmic male sterility locus in Brassica cybrids is determined post-transcriptionally by its 3' region

Abstract

We have investigated the control of the expression of three different configurations of the mitochondrial gene orf138, whose expression is correlated with Ogura cytoplasmic male-sterility in rapeseed cybrids. These configurations, termed Nco2.5/13S, Nco2.7/13F and Bam4.8/18S, specific to the 13S (sterile), 13F (fertile) and 18S (sterile) cybrids respectively, have the same 5' regions but different 3' regions. The orf138 transcript from Bam4.8/18S is 10-fold more abundant than the one from Nco2.5/13S, while no orf138 transcript from Nco2.7/13F accumulates. However, transcriptional activity measurements show that the rate of transcription is equivalent for the three configurations. These results strongly suggest that the steady-state level of mRNA from the orf138 locus is determined post-transcriptionally, most likely by its 3' region. To determine the role of these 3' regions, we have established an in vitro decay and processing system. In the presence of rapeseed mitochondrial lysate, synthetic RNAs corresponding to the 3' region of the Nco2.7/13F transcript are, as expected, less stable than RNAs corresponding to the 3' regions of the Nco2.5/13S and Bam4.8/18S transcripts. We have also observed in vitro processing of synthetic RNAs at the sites corresponding to the 3' ends of the natural mRNAs from Nco2.5/13S and Bam4.8/18S. Further analysis of the role of these 3' regions in in vitro RNA stability should help us to better understand post-transcriptional control in plant mitochondria.