scholarly journals Expanding the Genetic Code ofCaenorhabditis elegansUsing Bacterial Aminoacyl-tRNA Synthetase/tRNA Pairs

2012 ◽  
Vol 7 (7) ◽  
pp. 1292-1302 ◽  
Author(s):  
Angela R. Parrish ◽  
Xingyu She ◽  
Zheng Xiang ◽  
Irene Coin ◽  
Zhouxin Shen ◽  
...  
Keyword(s):  
Genetic Code ◽  
Trna Synthetase ◽  
Aminoacyl Trna Synthetase

scholarly journals Pyrrolysyl-tRNA Synthetase, an Aminoacyl-tRNA Synthetase for Genetic Code Expansion

2016 ◽  
Vol 89 (2) ◽  
Author(s):  
Ana Crnković ◽  
Tateki Suzuki ◽  
Dieter Söll ◽  
Noah M. Reynolds
Keyword(s):  
Genetic Code ◽  
Trna Synthetase ◽  
Aminoacyl Trna Synthetase ◽  
Genetic Code Expansion

Molecular Evolution of Aminoacyl tRNA Synthetases and Origin of Universal Genetic Code

1997 ◽  
Vol 161 ◽  
pp. 483-489
Author(s):  
Masahiro Ishigami ◽  
Hideshi Ihara ◽  
Hiroyuki Shinoda
Keyword(s):  
Genetic Code ◽  
Trna Synthetase ◽  
Code System ◽  
Aminoacyl Trna Synthetase ◽  
Synthesis Mechanism ◽  
Primitive Earth ◽  
Universal Genetic Code ◽  
Trna Synthetases ◽  
Aminoacyl Trna Synthetases ◽  
Living Things

AbstractIt is thought that living things first appeared on the primitive earth 35 hundred million years ago. At that time, a primitive protein synthesis mechanism was thought to have been established, a genetic code system evolved, and a universal genetic code system developed. Aminoacyl tRNA synthetase must have evolved with the genetic code system. The aim of the present study is to clarify the evolution of aminoacyl tRNA synthetase and the process and era of the establishment of the universal genetic code system.

scholarly journals An Expanded Genetic Code in Candida albicans To Study Protein-Protein Interactions In Vivo

2013 ◽  
Vol 12 (6) ◽  
pp. 816-827 ◽  
Author(s):  
Silke Palzer ◽  
Yannick Bantel ◽  
Franziska Kazenwadel ◽  
Michael Berg ◽  
Steffen Rupp ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Candida Albicans ◽  
Amino Acid ◽  
Genetic Code ◽  
Molecular Interactions ◽  
Trna Synthetase ◽  
Aminoacyl Trna Synthetase ◽  
Content Type ◽  
Suppressor Trna

ABSTRACT For novel insights into the pathogenicity of Candida albicans , studies on molecular interactions of central virulence factors are crucial. Since methods for the analysis of direct molecular interactions of proteins in vivo are scarce, we expanded the genetic code of C. albicans with the synthetic photo-cross-linking amino acid p -azido- l -phenylalanine (AzF). Interacting molecules in close proximity of this unnatural amino acid can be covalently linked by UV-induced photo-cross-link, which makes unknown interacting molecules available for downstream identification. Therefore, we applied an aminoacyl-tRNA synthetase and a suppressor tRNA pair ( Ec TyrtRNA CUA ) derived from Escherichia coli , which was previously reported to be orthogonal in Saccharomyces cerevisiae . We further optimized the aminoacyl-tRNA synthetase for AzF (AzF-RS) and Ec TyrtRNA CUA for C. albicans and identified one AzF-RS with highest charging efficiency. Accordingly, incorporation of AzF into selected model proteins such as Tsa1p or Tup1p could be considerably enhanced. Immunologic detection of C-terminally tagged Tsa1p and Tup1p upon UV irradiation in a strain background containing suppressor tRNA and optimized AzF-RS revealed not only the mutant monomeric forms of these proteins but also higher-molecular-weight complexes, strictly depending on the specific position of incorporated AzF and UV excitation. By Western blotting and tandem mass spectrometry, we could identify these higher-molecular-weight complexes as homodimers consisting of one mutant monomer and a differently tagged, wild-type version of Tsa1p or Tup1p, respectively, demonstrating that expanding the genetic code of C. albicans with the unnatural photo-cross-linker amino acid AzF and applying it for in vivo binary protein interaction analyses is feasible.

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