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PMID: 18782833 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The human mitochondrial ribosome recycling factor is essential for cell viability.

Nucleic acids research ·Vol. 36 ·No. 18 ·2008-10-00 ·Pages 5787-99

Rorbach J, Richter R, Wessels HJ, Wydro M, Pekalski M, Farhoud M, Kühl I, Gaisne M, Bonnefoy N, Smeitink JA, Lightowlers RN, Chrzanowska-Lightowlers ZM

Abstract

The molecular mechanism of human mitochondrial translation has yet to be fully described. We are particularly interested in understanding the process of translational termination and ribosome recycling in the mitochondrion. Several candidates have been implicated, for which subcellular localization and characterization have not been reported. Here, we show that the putative mitochondrial recycling factor, mtRRF, is indeed a mitochondrial protein. Expression of human mtRRF in fission yeast devoid of endogenous mitochondrial recycling factor suppresses the respiratory phenotype. Further, human mtRRF is able to associate with Escherichia coli ribosomes in vitro and can associate with mitoribosomes in vivo. Depletion of mtRRF in human cell lines is lethal, initially causing profound mitochondrial dysmorphism, aggregation of mitoribosomes, elevated mitochondrial superoxide production and eventual loss of OXPHOS complexes. Finally, mtRRF was shown to co-immunoprecipitate a large number of mitoribosomal proteins attached to other mitochondrial proteins, including putative members of the mitochondrial nucleoid.

MeSH Terms
Amino Acid Sequence Cell Death Cell Line Cell Proliferation Cell Respiration Cell Survival Escherichia coli/genetics HeLa Cells Humans Immunoprecipitation Mitochondria/genetics,ultrastructure Mitochondrial Proteins/antagonists & inhibitors,genetics,physiology Molecular Sequence Data Protein Biosynthesis Reactive Oxygen Species/metabolism Ribosomal Proteins/antagonists & inhibitors,genetics,physiology Ribosomes/metabolism Schizosaccharomyces/genetics,metabolism Schizosaccharomyces pombe Proteins/genetics
Chemicals
Mitochondrial Proteins Reactive Oxygen Species Ribosomal Proteins Schizosaccharomyces pombe Proteins ribosome releasing factor
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Rorbach Joanna
Mitochondrial Research Group, Institute of Cellular Medicine, Medical School, Newcastle University, Framlington Place, Newcastle upon Tyne, NE2 4HH, UK.
Richter Ricarda
Wessels Hans J
Wydro Mateusz
Pekalski Marcin
Farhoud Murtada
Kühl Inge
Gaisne Mauricette
Bonnefoy Nathalie
Smeitink Jan A
Lightowlers Robert N
Chrzanowska-Lightowlers Zofia M A
References (40)
40 references, click to expand
  1. Post-transcriptional regulation of the steady-state levels of mitochondrial tRNAs in HeLa cells.
    J Biol Chem. 1993 May 15;268(14):10228-37 PMID: 7683672
  2. Splitting of the posttermination ribosome into subunits by the concerted action of RRF and EF-G.
    Mol Cell. 2005 Jun 10;18(6):675-86 PMID: 15949442
  3. Dual functions of ribosome recycling factor in protein biosynthesis: disassembling the termination complex and preventing translational errors.
    Biochimie. 1996;78(11-12):959-69 PMID: 9150873
  4. The termination of translation.
    Curr Opin Struct Biol. 2008 Feb;18(1):70-7 PMID: 18206363
  5. Sequence and organization of the human mitochondrial genome.
    Nature. 1981 Apr 9;290(5806):457-65 PMID: 7219534
  6. The large subunit of the mammalian mitochondrial ribosome. Analysis of the complement of ribosomal proteins present.
    J Biol Chem. 2001 Nov 23;276(47):43958-69 PMID: 11551941
  7. Sequence of steps in ribosome recycling as defined by kinetic analysis.
    Mol Cell. 2005 May 13;18(4):403-12 PMID: 15893724
  8. Blue Native electrophoresis to study mitochondrial and other protein complexes.
    Methods. 2002 Apr;26(4):327-34 PMID: 12054923
  9. Progression of the ribosome recycling factor through the ribosome dissociates the two ribosomal subunits.
    Mol Cell. 2007 Jul 20;27(2):250-61 PMID: 17643374
  10. Identification of mammalian mitochondrial translational initiation factor 3 and examination of its role in initiation complex formation with natural mRNAs.
    J Biol Chem. 2002 Sep 20;277(38):35541-9 PMID: 12095986
  11. Ribosome recycling factor (ribosome releasing factor) is essential for bacterial growth.
    Proc Natl Acad Sci U S A. 1994 May 10;91(10):4249-53 PMID: 8183897
  12. Temperature-sensitive mutation in yeast mitochondrial ribosome recycling factor (RRF).
    Nucleic Acids Res. 2003 Jul 15;31(14):4218-26 PMID: 12853640
  13. Identification and cloning of human mitochondrial translational release factor 1 and the ribosome recycling factor.
    Biochim Biophys Acta. 1998 Nov 26;1443(1-2):245-50 PMID: 9838146
  14. Evolutionary tinkering with mitochondrial nucleoids.
    Trends Cell Biol. 2007 Dec;17(12):586-92 PMID: 17981466
  15. The small subunit of the mammalian mitochondrial ribosome. Identification of the full complement of ribosomal proteins present.
    J Biol Chem. 2001 Jun 1;276(22):19363-74 PMID: 11279123
  16. Human mitochondrial DNA nucleoids are linked to protein folding machinery and metabolic enzymes at the mitochondrial inner membrane.
    J Biol Chem. 2006 Sep 1;281(35):25791-802 PMID: 16825194
  17. Specific interaction between the ribosome recycling factor and the elongation factor G from Mycobacterium tuberculosis mediates peptidyl-tRNA release and ribosome recycling in Escherichia coli.
    EMBO J. 2001 Jun 1;20(11):2977-86 PMID: 11387230
  18. Bovine mitochondrial protein synthesis elongation factors. Identification and initial characterization of an elongation factor Tu-elongation factor Ts complex.
    J Biol Chem. 1989 Nov 15;264(32):19125-31 PMID: 2808417
  19. Protein synthesis in mitochondria.
    Mol Biol Rep. 1994 May;19(3):183-94 PMID: 7969106
  20. Elongation factor G participates in ribosome disassembly by interacting with ribosome recycling factor at their tRNA-mimicry domains.
    Mol Cell. 2002 Jun;9(6):1263-72 PMID: 12086623
  21. Studying mitochondria in an attractive model: Schizosaccharomyces pombe.
    Methods Mol Biol. 2007;372:91-105 PMID: 18314720
  22. Crystal structure of the ribosome recycling factor bound to the ribosome.
    Nat Struct Mol Biol. 2007 Aug;14(8):733-7 PMID: 17660830
  23. Human mitochondrial ribosomal protein MRPL12 interacts directly with mitochondrial RNA polymerase to modulate mitochondrial gene expression.
    J Biol Chem. 2007 Apr 27;282(17):12610-8 PMID: 17337445
  24. Purification and characterization of elongation factor G from bovine liver mitochondria.
    J Biol Chem. 1990 Dec 5;265(34):21000-4 PMID: 2250005
  25. mtRF1a is a human mitochondrial translation release factor decoding the major termination codons UAA and UAG.
    Mol Cell. 2007 Sep 7;27(5):745-57 PMID: 17803939
  26. Expression of a recoded nuclear gene inserted into yeast mitochondrial DNA is limited by mRNA-specific translational activation.
    Proc Natl Acad Sci U S A. 1996 May 28;93(11):5253-7 PMID: 8643562
  27. Mitochondrial translation: elongation factor tu is essential in fission yeast and depends on an exchange factor conserved in humans but not in budding yeast.
    Genetics. 2005 Apr;169(4):1891-901 PMID: 15695360
  28. Initiation and beyond: multiple functions of the human mitochondrial transcription machinery.
    Mol Cell. 2006 Dec 28;24(6):813-25 PMID: 17189185
  29. Transcription and RNA-processing in fission yeast mitochondria.
    RNA. 2005 May;11(5):785-95 PMID: 15811919
  30. In vivo labeling and analysis of human mitochondrial translation products.
    Methods Enzymol. 1996;264:197-211 PMID: 8965693
  31. Interaction of ribosome recycling factor and elongation factor EF-G with E. coli ribosomes studied by the surface plasmon resonance technique.
    Genes Cells. 2000 Dec;5(12):953-63 PMID: 11168582
  32. The COX18 gene, involved in mitochondrial biogenesis, is functionally conserved and tightly regulated in humans and fission yeast.
    FEMS Yeast Res. 2006 Sep;6(6):869-82 PMID: 16911509
  33. The layered structure of human mitochondrial DNA nucleoids.
    J Biol Chem. 2008 Feb 8;283(6):3665-3675 PMID: 18063578
  34. Assembly of mitochondrial cytochrome c-oxidase, a complicated and highly regulated cellular process.
    Am J Physiol Cell Physiol. 2006 Dec;291(6):C1129-47 PMID: 16760263
  35. Specific polar localization of ribosomes in Bacillus subtilis depends on active transcription.
    EMBO Rep. 2001 Aug;2(8):685-9 PMID: 11463749
  36. The ribosome-recycling step: consensus or controversy?
    Trends Biochem Sci. 2006 Mar;31(3):143-9 PMID: 16487710
  37. Visualization of ribosome-recycling factor on the Escherichia coli 70S ribosome: functional implications.
    Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):8900-5 PMID: 15178758
  38. A regulatory factor, Fil1p, involved in derepression of the isocitrate lyase gene in Saccharomyces cerevisiae--a possible mitochondrial protein necessary for protein synthesis in mitochondria.
    Eur J Biochem. 1998 Aug 15;256(1):212-20 PMID: 9746366
  39. The AAA+ protein ATAD3 has displacement loop binding properties and is involved in mitochondrial nucleoid organization.
    J Cell Biol. 2007 Jan 15;176(2):141-6 PMID: 17210950
  40. Cloning and sequencing of a human cDNA coding for dihydroorotate dehydrogenase by complementation of the corresponding yeast mutant.
    Gene. 1992 Nov 16;121(2):393-6 PMID: 1446837
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkn576
Published
2008-10-00
Epub
2008-00-09
Pages
5787-99
Language
English
Region
England
NLM ID
0411011
PMCID
PMC2566884
Subset
IM
Grants
Wellcome Trust · 074454 · United Kingdom
Biotechnology and Biological Sciences Research Council · BB/F011520/1 · United Kingdom
Wellcome Trust · 074454/Z/04/Z · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/gkn576 PMC Full Text View journal details
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