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Vol. 16, Issue 9, 4172-4182, September 2005
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Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada
Submitted March 7, 2005;
Revised June 21, 2005;
Accepted June 22, 2005
Monitoring Editor: Thomas Fox
The TOM (translocase of the outer mitochondrial membrane) complex of the outer mitochondrial membrane is required for the import of proteins into the organelle. The core TOM complex contains five proteins, including three small components Tom7, Tom6, and Tom5. We have created single and double mutants of all combinations of the three small Tom proteins of Neurospora crassa. Analysis of the mutants revealed that Tom6 plays a major role in TOM complex stability, whereas Tom7 has a lesser role. Mutants lacking both Tom6 and Tom7 have an extremely labile TOM complex and are the only class of mutant to exhibit an altered growth phenotype. Although single mutants lacking N. crassa Tom5 have no apparent TOM complex abnormalities, studies of double mutants lacking Tom5 suggest that it also has a minor role in maintaining TOM complex stability. Our inability to isolate triple mutants supports the idea that the three proteins have overlapping functions. Mitochondria lacking either Tom6 or Tom7 are differentially affected in their ability to import different precursor proteins into the organelle, suggesting that they may play roles in the sorting of proteins to different mitochondrial subcompartments. Newly imported Tom40 was readily assembled into the TOM complex in mitochondria lacking any of the small Tom proteins.
Abbreviations used: AAC, ADP-ATP carrier protein; BNGE, blue native gel electrophoresis; DIG, digitonin; DDM, n-dodecyl-
-D-maltoside; F1
, beta subunit of the F1 ATP synthase; RIP, repeat induced point mutation; TOM, translocase of the outer mitochondrial membrane.
* Present address: Department of Agricultural, Food and Nutritional Science, 4-10 Agriculture/Forestry Centre, University of Alberta, Edmonton, Alberta T6G 2P5, Canada.
Address correspondence to: Frank E. Nargang (frank.nargang{at}ualberta.ca).
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