In yeast, the 3' untranslated region or the presequence of ATM1 is required for the exclusive localization of its mRNA to the vicinity of mitochondria.
We isolated mitochondria from Saccharomyces cerevisiae to selectively study polysomes bound to the mitochondrial surface. The distribution of several mRNAs coding for mitochondrial proteins was examined in free and mitochondrion-bound polysomes. Some mRNAs exclusively localize to mitochondrion-bound polysomes, such as the ones coding for Atm1p, Cox10p, Tim44p, Atp2p, and Cot1p. In contrast, mRNAs encoding Cox6p, Cox5a, Aac1p, and Mir1p are found enriched in free cytoplasmic polysome fractions. Aac1p and Mir1p are transporters that lack cleavable presequences. Sequences required for mRNA asymmetric subcellular distribution were determined by analyzing the localization of reporter mRNAs containing the presequence coding region and/or the 3'-untranslated region (3'UTR) of ATM1, a gene encoding an ABC transporter of the mitochondrial inner membrane. Biochemical analyses of mitochondrion-bound polysomes and direct visualization of RNA localization in living yeast cells allowed us to demonstrate that either the presequence coding region or the 3'UTR of ATM1 is sufficient to allow the reporter mRNA to localize to the vicinity of the mitochondrion, independently of its translation. These data demonstrate that mRNA localization is one of the mechanisms used, in yeast, for segregating mitochondrial proteins.
Corral-Debrinski
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M
Laboratoire de Génétique Moléculaire, UMR CNRS 8541, Ecole Normale Supérieure, 75230 Paris, France. corral@biologie.ens.fr
Blugeon
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C
Jacq
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D020413:3' Untranslated Regions; D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism*; D015152:Blotting, Northern; D003593:Cytoplasm / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism; D017353:Gene Deletion; D017930:Genes, Reporter; D005947:Glucose / Q000378:metabolism; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D015636:Magnesium Chloride / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins
D020413:3' Untranslated Regions; C091561:ATM1 protein, S cerevisiae; D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D008164:Luminescent Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D015636:Magnesium Chloride; D049452:Green Fluorescent Proteins; D004492:Edetic Acid; D005947:Glucose; D005690:Galactose