Mitochondrial Form of a tRNA Synthetase Can Be Made Bifunctional by Manipulating Its Leader Peptide

Wang, Chien Chia; Chang, Kuang Jung; Hsieh, Chia‐Jung; Schimmel, Paul

doi:10.1021/bi025964c

Cited by 29 publications

(37 citation statements)

References 21 publications

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“…As previously shown, VAS1c, which has its mitochondrial targeting peptide (residues 1-46) deleted, encodes only the cytoplasmic form of valyl-tRNA synthetase and thus, cannot rescue the mitochondrial defect of the VAS1 knockout strain CW1 (22). Substitution of the native VAS1 promoter in VAS1c with the GRS1 presequence enabled the fusion to provide both the cytoplasmic and mitochondrial activities (preGRS1-VAS1c in Fig.…”

Section: Initiation Of the Mitochondrial Isoform From A Native Non-aumentioning

confidence: 87%

“…Although the leader peptide is required for mitochondrial import of this enzyme under normal conditions, overexpression of a leaderless form enabled it to overcome this requisite (35). In contrast to these observations, although VAS1 also encodes two distinct protein isoforms through alternative transcription and translation, the cytoplasmic form cannot complement its mitochondrial function even when it is overexpressed (22).…”

Section: Discussionmentioning

confidence: 98%

“…ADH/GRS1⌬2-12 was constructed as previously described (17). For construction of ADH/GRS1, the open reading frame (extending from base pair Ϫ3 to the stop codon) of GRS1 was amplified by PCR as an EagI-SalI fragment and cloned in a high-copy number plasmid pADH1 (22). Cloning of ADH/GRS1 Ϫ88 ⌬2-12 (extending from base pair Ϫ88 to the stop codon but lacking codons 2-12) in pADH1 followed a similar protocol.…”

Section: Methodsmentioning

confidence: 99%

“…Construction of wild-type VAS1 (designated as wt VAS1) and VAS1c has been described previously (22). For construction of preGRS1-VAS1c, a DNA sequence containing base pairs Ϫ300 to Ϫ1 relative to ATG 1 of GRS1 was amplified by PCR and used to substitute for the corresponding EagI-NdeI fragment in VAS1c, yielding preGRS1-VAS1c.…”

Section: Methodsmentioning

confidence: 99%

“…We next tested whether the short form of GlyRS can be converted into a mitochondrial enzyme by a well characterized mitochondrial targeting signal (22). Fig.…”

Section: Initiation Of the Mitochondrial Isoform From A Native Non-aumentioning

confidence: 99%

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Translation Initiation from a Naturally Occurring Non-AUG Codon in Saccharomyces cerevisiae

Chang

Wang

2004

Journal of Biological Chemistry

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117

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Although previous studies have already shown that both cytoplasmic and mitochondrial activities of glycyltRNA synthetase are provided by a single gene, GRS1, in the yeast Saccharomyces cerevisiae, the mechanism by which this occurs remains unclear. Evidence presented here indicates that this bifunctional property is actually a result of two distinct translational products alternatively generated from a single transcript of this gene. Except for an amino-terminal 23-amino acid extension, these two isoforms have the same polypeptide sequence and function exclusively in their respective compartments under normal conditions. Reporter gene assays further suggest that this leader peptide can function independently as a mitochondrial targeting signal and plays the major role in the subcellular localization of the isoforms. Additionally, whereas the short protein is translationally initiated from a traditional AUG triplet, the longer isoform is generated from an upstream inframe UUG codon. To our knowledge, GRS1 appears to be the first example in the yeast wherein a functional protein isoform is initiated from a naturally occurring non-AUG codon. The results suggest that non-AUG initiation might be a mechanism existing throughout all kingdoms.

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Section: Initiation Of the Mitochondrial Isoform From A Native Non-aumentioning

confidence: 87%

Section: Discussionmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…We next tested whether the short form of GlyRS can be converted into a mitochondrial enzyme by a well characterized mitochondrial targeting signal (22). Fig.…”

Section: Initiation Of the Mitochondrial Isoform From A Native Non-aumentioning

confidence: 99%

See 3 more Smart Citations

Translation Initiation from a Naturally Occurring Non-AUG Codon in Saccharomyces cerevisiae

Chang

Wang

2004

Journal of Biological Chemistry

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117

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A combination of two novel VARS2 variants causes a mitochondrial disorder associated with failure to thrive and pulmonary hypertension

et al. 2019

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Dual targeting of mitochondrial proteins: Mechanism, regulation and function

Yogev

Pines

2011

Biochimica et Biophysica Acta (BBA) - Biomembranes

174

160

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One solution found in evolution to increase the number of cellular functions, without increasing the number of genes, is distribution of single gene products to more than one cellular compartment. It is well documented that in eukaryotic cells, molecules of one protein can be located in several subcellular locations, a phenomenon termed dual targeting, dual localization, or dual distribution. The differently localized proteins are coined in this review "echoforms" indicating repetitious forms of the same protein (echo in Greek denotes repetition) distinctly placed in the cell. This term replaces the term to "isoproteins" or "isoenzymes" which are reserved for proteins with the same activity but different amino acid sequences. Echoforms are identical or nearly identical, even though, as referred to in this review may, in some cases, surprisingly have a totally different function in the different compartments. With regard to mitochondria, our operational definition of dual targeted proteins refers to situations in which one of the echoforms is translocated through/into a mitochondrial membrane. In this review we ask how, when and why mitochondrial proteins are dual localized in the cell. We describe mechanisms of dual targeting of proteins between mitochondria and other compartments of the eukaryotic cell. In particular, we have paid attention to situations in which dual localization is regulated in time, location or function. In addition, we have attempted to provide a broader view concerning the phenomenon of dual localization of proteins by looking at mechanisms that are beyond our simple definition of dual targeting. This article is part of a Special Issue entitled Protein translocation across or insertion into membranes.

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Mitochondrial Form of a tRNA Synthetase Can Be Made Bifunctional by Manipulating Its Leader Peptide

Cited by 29 publications

References 21 publications

Translation Initiation from a Naturally Occurring Non-AUG Codon in Saccharomyces cerevisiae

Translation Initiation from a Naturally Occurring Non-AUG Codon in Saccharomyces cerevisiae

A combination of two novel VARS2 variants causes a mitochondrial disorder associated with failure to thrive and pulmonary hypertension

Dual targeting of mitochondrial proteins: Mechanism, regulation and function

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