Identification of nuclear encoded genes involved in 5' end processing of mitochondrial mRNA in Arabidopsis thaliana

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In the proposed project genes involved in 5 end processing of plant mitochondrial transcripts will be identified by a forward genetic approach. The groundwork for this project is the analysis of major mitochondrial mRNA ends in three different ecotypes of Arabidopsis thaliana. This investigation revealed ecotype-specific 5’ mRNA polymorphisms for the mitochondrial genes nad9, nad4, ccmB and rpl5. Analysis of corresponding F1 hybrids assigned these polymorphisms to different nuclear encoded genes. Now the naturally occurring genetic variation in the model species A. thaliana will be used for map-based cloning of these nuclear genes encoding mitochondrial RNA processing factors (RPF). The highly valuable system of natural variation is presently the sole alternative to the very limited cytoplasmic male sterility/restorer of fertility system, which is not found for A. thaliana. To identify further ecotype-specific variations a comprehensive descriptive analysis of 5’ processing of mRNA derived from gene encoding subunits of complex I (nad genes) will be initiated in parallel. This investigation will yield more definite information about the processing events and of RNA intermediates.