Yeast Nfs1p Is Involved in Thio-modification of Both Mitochondrial and Cytoplasmic tRNAs

Nakai, Y.; Umeda, N.; Suzuki, Tsutomu; Nakai, Masaaki; Hayashi, Hideyuki; Watanabe, Kimitsuna; Kagamiyama, Hiroyuki

doi:10.1074/jbc.m312448200

Cited by 113 publications

(117 citation statements)

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“…tRNA Extraction and Gel Retardation Analysis of the Thiomodification of tRNA-The total tRNAs of the yeast cells were prepared as described previously (3). The plant seedlings were grown on Murashige and Skoog plates for 3 weeks, and tRNAs were extracted using the PureLink miRNA Isolation kit (Invitrogen).…”

Section: Plant and Yeast Strains Plasmids Media And Growthmentioning

confidence: 99%

“…The molecular mechanisms for tRNA thio-modification in eukaryotic cells have primarily been studied using yeast, especially Saccharomyces cerevisiae. In the yeast S. cerevisiae, cytosolic and mitochondrial tRNAs are thio-modified by partially overlapping multistep mechanisms in which several cytosolic and mitochondrial proteins participate (3)(4)(5)(6)(7). A common initial step for these thiomodifications involves the mitochondrial cysteine desulfurase Nfs1, which cleaves sulfur from its substrate cysteine to form the enzyme-bound cysteinyl persulfide intermediate (3).…”

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confidence: 99%

“…In the yeast S. cerevisiae, cytosolic and mitochondrial tRNAs are thio-modified by partially overlapping multistep mechanisms in which several cytosolic and mitochondrial proteins participate (3)(4)(5)(6)(7). A common initial step for these thiomodifications involves the mitochondrial cysteine desulfurase Nfs1, which cleaves sulfur from its substrate cysteine to form the enzyme-bound cysteinyl persulfide intermediate (3). Sulfur is then incorporated into either cytosolic or mitochondrial substrate tRNAs through distinct sulfur-transferring systems (4).…”

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confidence: 99%

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Arabidopsis Molybdopterin Biosynthesis Protein Cnx5 Collaborates with the Ubiquitin-like Protein Urm11 in the Thio-modification of tRNA

Nakai

Harada

Hashiguchi

et al. 2012

Journal of Biological Chemistry

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Background:The plant proteins responsible for tRNA thio-modification are unclear. Results: Ubiquitin-like protein Urm11 and the molybdopterin biosynthesis protein Cnx5 participate together in tRNA thiomodification in Arabidopsis. Conclusion: Cnx5 is a ubiquitin-activating enzyme-like protein that functions in two different metabolic processes with two distinct ubiquitin-like partners. Significance: Cnx5-Urm11 ortholog pair-mediated thio-modification of tRNA is ubiquitous in most eukaryotes, including plants.

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Section: Plant and Yeast Strains Plasmids Media And Growthmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Arabidopsis Molybdopterin Biosynthesis Protein Cnx5 Collaborates with the Ubiquitin-like Protein Urm11 in the Thio-modification of tRNA

Nakai

Harada

Hashiguchi

et al. 2012

Journal of Biological Chemistry

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“…The involvement of components in the ISC biogenesis pathway for thio modification of eukaryotic tRNAs has so far mainly been analyzed in yeast (15,16). In the present study we have extended this analysis to the parasitic protozoa T. brucei.…”

Section: Discussionmentioning

confidence: 76%

“…Moreover, the same appears to be the case for the corresponding mitochondrial encoded tRNAs (14). Recent work in the yeast Saccharomyces cerevisiae has shown that thiolation of tRNAs depends on components of the Fe/S cluster (ISC) biogenesis pathway (15)(16)(17). ISCs of iron-sulfur proteins are essential for a wide variety of cellular processes, and their assembly requires complex and interconnected machineries that are localized in both mitochondria and the cytosol (17).…”

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Thiolated tRNAs of Trypanosoma brucei Are Imported into Mitochondria and Dethiolated after Import

Bruske

Sendfeld

Schneider

2009

Journal of Biological Chemistry

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All mitochondrial tRNAs in Trypanosoma brucei derive from cytosolic tRNAs that are in part imported into mitochondria. Some trypanosomal tRNAs are thiolated in a compartment-specific manner. We have identified three proteins required for the thio modification of cytosolic tRNA Gln , tRNA Glu , and tRNA Lys . RNA interference-mediated ablation of these proteins results in the cytosolic accumulation non-thio-modified tRNAs but does not increase their import. Moreover, in vitro import experiments showed that both thio-modified and non-thio-modified tRNA Glu can efficiently be imported into mitochondria. These results indicate that unlike previously suggested the cytosolspecific thio modifications do not function as antideterminants for mitochondrial tRNA import. Consistent with these results we showed by using inducible expression of a tagged tRNA Glu that it is mainly the thiolated form that is imported in vivo. Unexpectedly, the imported tRNA becomes dethiolated after import, which explains why the non-thiolated form is enriched in mitochondria. Finally, we have identified two genes required for thiolation of imported tRNA Trp whose wobble nucleotide is subject to mitochondrial C to U editing. Interestingly, downregulation of thiolation resulted in an increase of edited tRNA Trp but did not affect growth.Most protozoa, many fungi, plants, and a few animals lack a variable number of mitochondrial tRNA genes. It has been shown in these organisms that the missing genes are compensated for by import of a small fraction of the corresponding cytosolic tRNAs (1, 2). Among all organisms that import tRNAs trypanosomatids such as Trypanosoma brucei and Leishmania are extreme in that their mitochondrial genomes have lost all tRNA genes. Trypanosomatids therefore need to import the entire set of mitochondrial tRNAs. As a consequence all mitochondrial tRNAs in trypanosomatids derive from eukaryotic-type cytosolic tRNAs that need to function in the context of the bacterial-type translation system of mitochondria (3). Two tRNAs, the initiator tRNA Met and the tRNA Sec , are cytosol-specific. It has been shown that in T. brucei the tRNA import specificity is mediated by binding to cytosolic translation elongation factor 1a (4). The fact that initiator tRNA Met and tRNA Sec do not bind to elongation factor 1a therefore explains their exclusive cytosolic localization.The extent of mitochondrial localization of different trypanosomal tRNAs varies by at least an order of magnitude (5, 6). The same is true for other organisms that import tRNAs (7). Cells therefore need to determine both the tRNA import specificity as well as the extent of mitochondrial localization of each imported tRNA species. How this is achieved is not known in any species.In trypanosomatids mitochondrial tRNAs and their cytosolic counterparts derive from the same nuclear genes. However, due to compartment-specific post-transcriptional nucleotide modifications, the cytosolic and the corresponding imported tRNAs are often physically different. Mitochondria-specifi...

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Eclipsed distribution: A phenomenon of dual targeting of protein and its significance

Regev‐Rudzki

Pines

2007

BioEssays

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One of the surprises from genome sequencing projects is the apparently small number of predicted genes in different eukaryotic cells, particularly human. One possible reason for this 'shortage' of genes is multiple distribution of proteins; a single protein is targeted to more than one subcellular compartment and consequently participates in different biochemical pathways and might have completely different functions. Indeed, in recent years, there have been reports on proteins that were found to be localized in cellular compartments other than those initially attributed to them. Furthermore, the phenomenon of highly uneven isoprotein distribution was recently observed and termed 'eclipsed distribution'. In these cases, the amount of one of the isoproteins, in one of the locations, is significantly minute and its detection by standard biochemical and visualization methods is masked by the presence of the dominant isoprotein. In fact, the minute amounts of eclipsed proteins can be essential. Since detecting eclipsed distribution is difficult, we assume that this phenomenon is probably more common than currently recorded. Hence, developing methods for localization and functional detection of eclipsed proteins is a challenge in cell biology research. Finally, eclipsed distribution may lead to cellular pathologies as has been suggested to occur in human disorders such as Prion diseases and Alzheimer. This review provides a short description of the eclipsed distribution phenomenon followed by an overview of protein distribution mechanisms, examples of eclipsed distribution and experimental approaches for revealing these elusive proteins.

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Yeast Nfs1p Is Involved in Thio-modification of Both Mitochondrial and Cytoplasmic tRNAs

Cited by 113 publications

References 46 publications

Arabidopsis Molybdopterin Biosynthesis Protein Cnx5 Collaborates with the Ubiquitin-like Protein Urm11 in the Thio-modification of tRNA

Arabidopsis Molybdopterin Biosynthesis Protein Cnx5 Collaborates with the Ubiquitin-like Protein Urm11 in the Thio-modification of tRNA

Thiolated tRNAs of Trypanosoma brucei Are Imported into Mitochondria and Dethiolated after Import

Eclipsed distribution: A phenomenon of dual targeting of protein and its significance

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