In vivo selection of lethal mutations reveals two functional domains in arginyl–tRNA synthetase

Geslain, Renaud; Martin, Franck; Delagoutte, B.; Cavarelli, Jean; Gangloff, Jean; Eriani, Gilbert

doi:10.1017/s1355838200992331

Cited by 18 publications

(24 citation statements)

References 52 publications

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“…For HisRS, Rhizopus delemar (R03G_01784 and RO3G_16958) and Phycomyces blakesleeanus (PHYBL_135135 and PHYBL_138952) likewise contain gene duplications. Similarly, S. cerevisiae has two ArgRS genes that arose from the whole-genome duplication: RRS1/YDR341C is essential, abundant, and inferred to be cytoplasmic (54) while MSK1/YHR091C has a mitochondrial localization sequence and MSR1 deletions have a petite phenotype (55), although both have been detected in mitochondria suggesting some residual dual-localization of the cytoplasmic enzyme (56). The second S. cerevisiae stress-responsive cytoplasmic copy of GlyRS also arose from the whole-genome duplication (57).…”

Section: Resultsmentioning

confidence: 99%

Start codon context controls translation initiation in the fungal kingdom

Wallace

Maufrais

Sales-Lee

et al. 2019

Preprint

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Eukaryotic protein synthesis initiates at a start codon defined by an AUG and its surrounding Kozak sequence context, but studies of S. cerevisiae suggest this context is of little importance in fungi. We tested this concept in two pathogenic Cryptococcus species by genome-wide mapping of translation and of mRNA 5’ and 3’ ends. We observed that upstream open reading frames (uORFs) are a major contributor to translation repression, that uORF use depends on the Kozak sequence context of its start codon, and that uORFs with strong contexts promote nonsense-mediated mRNA decay. Numerous Cryptococcus mRNAs encode predicted dual-localized proteins, including many aminoacyl-tRNA synthetases, in which a leaky AUG start codon is followed by a strong Kozak context in-frame AUG, separated by mitochondrial-targeting sequence. Further analysis shows that such dual-localization is also predicted to be common in Neurospora crassa. Kozak-controlled regulation is correlated with insertions in translational initiation factors in fidelity-determining regions that contact the initiator tRNA. Thus, start codon context is a signal that programs the expression and structures of proteins in fungi.

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Section: Resultsmentioning

confidence: 99%

Start codon context controls translation initiation in the fungal kingdom

Wallace

Maufrais

Sales-Lee

et al. 2019

Preprint

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“…The ␤ subunit should contain the second part of the active site and the helix bundle domain found in the class Ia synthetases (Arg-, Met-, Val-and IleRS). This domain is involved in tRNA-anticodon binding as shown by x-ray crystallography (30 -32) and functional studies (33,34). An additional domain located at the C-terminal end of the ␣ subunit, characteristic of the T. thermophilus and A. aeolicus enzymes corresponds to the split site for the A. aeolicus enzyme.…”

Section: Discussionmentioning

confidence: 97%

Leucyl-tRNA Synthetase Consisting of Two Subunits from Hyperthermophilic Bacteria Aquifex aeolicus

Xu¹,

Chen²,

Martin³

et al. 2002

Journal of Biological Chemistry

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In a hyperthermophilic bacterium, Aquifex aeolicus, leucyl-tRNA synthetase (LeuRS) consists of two nonidentical polypeptide subunits (␣ and ␤), different from the canonical LeuRS, which has a single polypeptide chain. By PCR, using genome DNA of A. aeolicus as a template, genes encoding the ␣ and ␤ subunits were amplified and cloned in Escherichia coli. The ␣ subunit could not be expressed stably in vivo, whereas the ␤ subunit was overproduced and purified by a simple procedure. The ␤ subunit was inactive in catalysis but was able to bind tRNA Leu . Interestingly, the heterodimer ␣␤-LeuRS could be overproduced in E. coli cells containing both genes and was purified to 95% homogeneity as a hybrid dimer. The kinetics of A. aeolicus LeuRS in presteady and steady states and cross-recognition of LeuRS and tRNA Leu from A. aeolicus and E. coli were studied. Magnesium concentration, pH value, and temperature aminoacylation optima were determined to be 12 mM, 7.8, and 70°C, respectively. Under optimal conditions, A. aeolicus ␣␤-LeuRS is stable up to 65°C.

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“…Novel probes for tRNA structure were validated (47) and methods adapted for use in tRNA research. They include, for example, cryo-EM (118), X-ray (119), genetic (120), proteomic (121), and bioinformatic (122,123) methods. Note the implementation of specialized databases (124)(125)(126).…”

Section: A Stimulating Start At Ibmpmentioning

confidence: 99%

History of tRNA research in strasbourg

Florentz

Giegé

2019

IUBMB Life

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The tRNA molecules, in addition to translating the genetic code into protein and defining the second genetic code via their aminoacylation by aminoacyl‐tRNA synthetases, act in many other cellular functions and dysfunctions. This article, illustrated by personal souvenirs, covers the history of ~60 years tRNA research in Strasbourg. Typical examples point up how the work in Strasbourg was a two‐way street, influenced by and at the same time influencing investigators outside of France. All along, research in Strasbourg has nurtured the structural and functional diversity of tRNA. It produced massive sequence and crystallographic data on tRNA and its partners, thereby leading to a deeper physicochemical understanding of tRNA architecture, dynamics, and identity. Moreover, it emphasized the role of nucleoside modifications and in the last two decades, highlighted tRNA idiosyncrasies in plants and organelles, together with cellular and health‐focused aspects. The tRNA field benefited from a rich local academic heritage and a strong support by both university and CNRS. Its broad interlinks to the worldwide community of tRNA researchers opens to an exciting future. © 2019 IUBMB Life, 2019 © 2019 IUBMB Life, 71(8):1066–1087, 2019

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In vivo selection of lethal mutations reveals two functional domains in arginyl–tRNA synthetase

Cited by 18 publications

References 52 publications

Start codon context controls translation initiation in the fungal kingdom

Start codon context controls translation initiation in the fungal kingdom

Leucyl-tRNA Synthetase Consisting of Two Subunits from Hyperthermophilic Bacteria Aquifex aeolicus

History of tRNA research in strasbourg

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