Yeast mitochondrial RNase P, RNase Z and the RNA degradosome are part of a stable supercomplex.
Initial steps in the synthesis of functional tRNAs require 5'- and 3'-processing of precursor tRNAs (pre-tRNAs), which in yeast mitochondria are achieved by two endonucleases, RNase P and RNase Z. In this study, using a combination of detergent-free Blue Native Gel Electrophoresis, proteomics and in vitro testing of pre-tRNA maturation, we reveal the physical association of these plus other mitochondrial activities in a large, stable complex of 136 proteins. It contains a total of seven proteins involved in RNA processing including RNase P and RNase Z, five out of six subunits of the mitochondrial RNA degradosome, components of the fatty acid synthesis pathway, translation, metabolism and protein folding. At the RNA level, there are the small and large rRNA subunits and RNase P RNA. Surprisingly, this complex is absent in an oar1Δ deletion mutant of the type II fatty acid synthesis pathway, supporting a recently published functional link between pre-tRNA processing and the FAS II pathway--apparently by integration into a large complex as we demonstrate here. Finally, the question of mt-RNase P localization within mitochondria was investigated, by GFP-tracing of a known protein subunit (Rpm2p). We find that about equal fractions of RNase P are soluble versus membrane-attached.
Robert-Cedergren Centre for Bioinformatics and Genomics, Department of Biochemistry, Université de Montréal, 2900 Edouard-Montpetit, Montreal, Quebec H3T 1J4, Canada.
Rachid
R
Daoud
Lise
L
Forget
B Franz
BF
Lang
D004722:Endoribonucleases / Q000032:analysis*; D064429:Fatty Acid Synthases / Q000235:genetics; D008928:Mitochondria / Q000201:enzymology / Q000235:genetics; D051336:Mitochondrial Membranes / Q000201:enzymology; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000302:isolation & purification; D021122:Protein Subunits / Q000032:analysis; D012323:RNA Processing, Post-Transcriptional; D012343:RNA, Transfer / Q000378:metabolism*; D043262:Ribonuclease P / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis; D017384:Sequence Deletion
D024101:Mitochondrial Proteins; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer; D064429:Fatty Acid Synthases; D004722:Endoribonucleases; C110250:RNase Z; C519164:RPM2 protein, S cerevisiae; D043262:Ribonuclease P