This article contains the following supporting material:
Only genes with a value above 9% are considered as having their mRNA translated in the vicinity of mitochondria (red). A minor intermediate class of 6 genes (scores of 4 to 6%) remains ambiguous and is thus included with the mRNAs translated on free cytoplasmic polysomes (scores of 0 to 3%) (blue).
An extension image J plug-in was developed (L. Jourdren et al in preparation) to automatically assess the minimum distance between an mRNA molecule, revealed by FISH, and the mitochondrio revealed as the fluorescence of the oligonucleotides hybridizing with the mt ribosomal RNA. A: 3-D reconstitution of mRNA and mitochondrial objects. B and C) Specific mRNA-linked fluorescence as a function of the distance to mitochondria.
After partial purification (Kellems et al. , 1975) the mitochondrial polysome mRNA content was calculated by a new method which took into account the different biases introduced by low purification yield and contamination (Garcia et al. , 2006). Specific oligonucleotides (Table S2) were designed to probe for the 107 mRNAs coding for mitochondrial complexes and for internal mitochondrial controls (COX1, COX2 mRNAs) and a contamination control (ACT1 mRNA). The table shows the results of three independent real-time PCR analyses of the 107 mRNAs. The MLR column indicates, when known, the MLR value previously determined by a global microarray approach (Marc et al. , 2002). These MLR values were ranked from 0 to 100, 100 representing a high probability for the corresponding mRNA to be in the vicinity of mitochondria. Note the general agreement with the RT-PCR analysis, which distinguishes two classes of mRNA (see Fig. S1): those linked to and those independent of mitochondria. The phylogenetic origin of the genes is indicated in red for prokaryotic origin and in white for eukaryotic origin. This information is from the psi-BLAST analyses available at SGD (Christie et al. , 2004).