Altering the Enantioselectivity of Tyrosyl-tRNA Synthetase by Insertion of a Stereospecific Editing Domain

Biochemistry
Charles J Richardson, E A First

Abstract

Translation of mRNAs by the ribosome is stereospecific, with only l-amino acids being incorporated into the nascent polypeptide chain. This stereospecificity results from the exclusion of d-amino acids at three steps during protein synthesis: (1) the aminoacylation of tRNA by aminoacyl-tRNA synthetases, (2) binding of aminoacyl-tRNAs to EF-Tu, and (3) recognition of aminoacyl-tRNAs by the ribosome. As a first step toward incorporating d-amino acids during protein synthesis, we have altered the enantioselectivity of tyrosyl-tRNA synthetase. This enzyme is unusual among aminoacyl-tRNA synthetases, as it can aminoacylate tRNA with d-tyrosine (albeit at a reduced rate compared to l-tyrosine). To change the enantioselectivity of tyrosyl-tRNA synthetase, we introduced the post-transfer editing domain from Pyrococcus horikoshii phenylalanyl-tRNA synthetase into the connective polypeptide 1 (CP1) domain of Geobacillus stearothermophilus tyrosyl-tRNA synthetase (henceforth designated TyrRS-FRSed). We show that the phenylalanyl-tRNA synthetase editing domain is stereospecific, hydrolyzing l-Tyr-tRNA(Tyr), but not d-Tyr-tRNA(Tyr). We further show that inserting the phenylalanyl-tRNA synthetase editing domain into the CP1 domain of tyrosyl...Continue Reading

References

Jan 1, 1978·Nucleic Acids Research·R S BrownG G Brownlee
Sep 1, 1992·Microbiological Reviews·H Jakubowski, E Goldman
Jun 14, 1991·Biochemical and Biophysical Research Communications·K FujimotoM Kobayashi
Aug 14, 1987·Biochemical and Biophysical Research Communications·R Shapira, C H Chou
Mar 20, 1987·Journal of Molecular Biology·P Brick, D M Blow
May 1, 1997·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·M Sela, E Zisman
Feb 9, 1999·EXS·D Ingrosso, A F Perna
Aug 5, 2000·The Journal of Biological Chemistry·J SoutourinaS Blanquet
Apr 21, 2001·Science·L WangP G Schultz
Jul 12, 2002·The EMBO Journal·Anna YaremchukStephen Cusack
May 29, 2003·Journal of the American Chemical Society·Larisa M DedkovaSidney M Hecht
Dec 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·Xiang-Lei YangLlúis Ribas de Pouplana
Jan 15, 2004·Nucleic Acids Research·Michael J Cavaluzzi, Philip N Borer
Oct 8, 2004·Journal of the American Chemical Society·Zhongping TanVirginia W Cornish
Mar 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·I E Segal
Sep 28, 2006·Proceedings of the National Academy of Sciences of the United States of America·Hiroshi M SasakiShigeyuki Yokoyama
Dec 21, 2006·Biochemistry·Larisa M DedkovaSidney M Hecht

❮ Previous
Next ❯

Citations

Apr 12, 2016·Biochemistry·Charles J Richardson, Eric Allen First
Oct 27, 2016·Methods : a Companion to Methods in Enzymology·Eric A First, Charles J Richardson
Dec 7, 2018·Nucleic Acids Research·Sergey V MelnikovYury S Polikanov
May 12, 2019·International Journal of Molecular Sciences·Ana CrnkovićDieter Söll
Mar 23, 2017·Biochimica Et Biophysica Acta. General Subjects·Noah M ReynoldsAna Crnković

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.