Biochemistry of Aminoacyl tRNA Synthetase and tRNAs and Their Engineering for Cell-Free and Synthetic Cell Applications

Ganesh, Ragunathan Bava; Maerkl, Sebastian J.

doi:10.3389/fbioe.2022.918659

Cited by 12 publications

(14 citation statements)

References 187 publications

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“…Using MALDI-TOF MS, we assigned the degradation products to 10-mer and 11-mer RNA strands, A 10 and A 11 , that had lost the terminal urea-modified nucleoside and the N 6 -methylcarbamoyl substituent, respectively. We also detected a 10-mer RNA strand with a terminal a 6 A nucleotide, A 9 -a 6 A. It seems that the nitrosation reaction provoked the partial degradation of the RNA strands.…”

Section: Methodsmentioning

confidence: 76%

“…To solve this chickenand-egg conundrum, we recently postulated that RNAamino acid conjugates could have been a starting point for evolving more complex systems. [11] We learned that RNA was capable of self-decorating with peptides with the help of non-canonical nucleosides, [12] that is, N 6 -methylated derivatives of glycine-and threonine-modified N 6 -carbamoyl adenosine, 1 a (g 6 A) [13] and 1 b (t 6 A). [14] In addition, 1 a-b are found in contemporary tRNAs in all three kingdoms of life [15,16] as potential relics of an early RNA world.…”

Section: Introductionmentioning

confidence: 99%

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Loading of Amino Acids onto RNA in a Putative RNA‐Peptide World

et al. 2023

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RNA is a molecule that can both store genetic information and perform catalytic reactions. This observed dualism places RNA into the limelight of concepts about the origin of life. The RNA world concept argues that life started from self-replicating RNA molecules, which evolved toward increasingly complex structures. Recently, we demonstrated that RNA, with the help of conserved non-canonical nucleosides, which are also putative relics of an early RNA world, had the ability to grow peptides covalently connected to RNA nucleobases, creating RNA-peptide chimeras. It is conceivable that such molecules, which combined the information-coding properties of RNA with the catalytic potential of amino acid side chains, were once the structures from which life emerged. Herein, we report prebiotic chemistry that enabled the loading of both nucleosides and RNAs with amino acids as the first step toward RNA-based peptide synthesis in a putative RNA-peptide world.

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Section: Methodsmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Loading of Amino Acids onto RNA in a Putative RNA‐Peptide World

et al. 2023

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“…This information (Table S2) was used to classify 20 aaRSs based on their requirements in a subsequent experiment. These different aaRS requirements for DNAP production were possibly caused by various factors, such as different numbers of each amino acid in the phi29 DNA polymerase (Table S3), different aminoacylation activities of each aaRS, and different aminoacyl-tRNA stabilities …”

Section: Resultsmentioning

confidence: 99%

In Vitro Transcription/Translation-Coupled DNA Replication through Partial Regeneration of 20 Aminoacyl-tRNA Synthetases

Hagino

Ichihashi

2023

ACS Synth. Biol.

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The in vitro reconstruction of life-like self-reproducing systems is a major challenge in in vitro synthetic biology. Self-reproduction requires regeneration of all molecules involved in DNA replication, transcription, and translation. This study demonstrated the continuous DNA replication and partial regeneration of major translation factors, 20 aminoacyl-tRNA synthetases (aaRS), in a reconstituted transcription/translation system (PURE system) for the first time. First, we replicated each DNA that encodes one of the 20 aaRSs through aaRS expression from the DNA by serial transfer experiments. Thereafter, we successively increased the number of aaRS genes and achieved simultaneous, continuous replication of DNA that encodes all 20 aaRSs, which comprised approximately half the number of protein factors in the PURE system, except for ribosomes, by employing dialyzed reaction and sequence optimization. This study provides a step-by-step methodology for continuous DNA replication with an increasing number of self-regenerative genes toward self-reproducing artificial systems.

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“…Er basiert auf informationstragender Boten-RNA (mRNA), dem Ribosom als katalytische Einheit und Transfer-RNAs (tRNAs), die spezifische Aminosäuren tragen und an die informationskodierenden Einheiten auf der mRNA binden. [1,2] Die Verknüpfung einer bestimmten Aminosäure mit der zugehörigen tRNA (Aminoacylierung) erfordert die Aktivierung der Aminosäure als Adenylat [3] und eine spezifische Aminoacyl-tRNA-Synthetase, [4][5][6] die die Aminosäure an das 3'-CCA-Ende der tRNA überträgt, indem sie die Bildung einer Esterbindung katalysiert. Obwohl tRNAs als hoch konservierte Strukturen gelten, [7][8][9][10] ist die Entstehung dieses komplexen Prozesses immer noch eine der wichtigsten ungelösten Fragen in der Forschung zum Ursprung des Lebens.…”

Section: Introductionunclassified

“…Um dieses Problem zu lösen, haben wir jüngst die These aufgestellt, dass RNA-Aminosäure-Konjugate ein Ausgangspunkt für die Entwicklung komplexerer Strukturen gewesen sein könnten. [11] Wir haben gelernt, dass RNA sich selbst mit Hilfe nicht-kanonischer Nukleoside, d. h. N 6 -methylierte Derivate von Glycin-und Threonin-modifiziertem N 6 -Carbamoyl-Adenosin, 1 a (g 6 A) [13] und 1 b (t 6 A), [14] mit Peptiden dekorieren kann. [12] Darüber hinaus sind 1 a-b als potentielle Relikte einer frühen RNA-Welt in den heutigen tRNAs in allen drei Reichen des Lebens zu finden.…”

Section: Introductionunclassified

Beladung von RNA mit Aminosäuren in einer putativen RNA‐Peptid‐Welt

et al. 2023

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Die RNA ist ein Molekül, das sowohl genetische Informationen speichern als auch katalytische Reaktionen ausführen kann. Dieser beobachtete Dualismus rückt die RNA ins Rampenlicht der Konzepte über den Ursprung des Lebens. Die RNA‐Welt‐Hypothese geht davon aus, dass das Leben aus selbst replizierenden RNA‐Molekülen entstand, die sich zu immer komplexeren Strukturen entwickelten. Vor kurzem haben wir gezeigt, dass RNA mit Hilfe erhaltener nicht‐kanonischer Nukleoside, die potenzielle Relikte einer frühen RNA‐Welt sind, in der Lage ist, Peptide zu bilden, die kovalent an Nukleobasen der RNA gebunden sind, wodurch RNA‐Peptid‐Chimären entstehen. Es ist denkbar, dass solche Moleküle, welche die informationskodierenden Eigenschaften der RNA mit dem katalytischen Potenzial von Aminosäureseitenketten kombinieren können, einst die Strukturen waren, aus denen das Leben entstand. In diesem Artikel berichten wir über präbiotische Chemie, die es ermöglicht sowohl Nukleoside als auch RNA mit Aminosäuren zu beladen, was einen ersten Schritt zu einer RNA‐basierten Peptidsynthese in einer mutmaßlichen RNA‐Peptid‐Welt darstellt.

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Biochemistry of Aminoacyl tRNA Synthetase and tRNAs and Their Engineering for Cell-Free and Synthetic Cell Applications

Cited by 12 publications

References 187 publications

Loading of Amino Acids onto RNA in a Putative RNA‐Peptide World

Loading of Amino Acids onto RNA in a Putative RNA‐Peptide World

In Vitro Transcription/Translation-Coupled DNA Replication through Partial Regeneration of 20 Aminoacyl-tRNA Synthetases

Beladung von RNA mit Aminosäuren in einer putativen RNA‐Peptid‐Welt

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