Changeability of individual domains of an aminoacyl-tRNA in polymerization by the ribosome.
The changeabilities of individual modules of aminoacyl-tRNAs are poorly understood, despite the relevance for evolution, translational accuracy and incorporation of unnatural amino acids (AAs). Here, we dissect the effect of successive changes in four domains of Ala-tRNA(3)(Ala) on translation in a purified system. Incorporating five AAs, not one, was necessary to reveal major effects on yields of peptide products. Omitting tRNA modifications had little affect, but anticodon mutations were very inhibitory. Surprisingly, changing the terminal CCA to CdCA was sometimes inhibitory and non-cognate AAs were sometimes compensatory. Results have implications for translational fidelity and engineering.
Gao
Rong
R
Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
Forster
Anthony C
AC
D000409:Alanine / Q000378:metabolism; D000412:Alanine-tRNA Ligase / Q000235:genetics / Q000378:metabolism*; D000926:Anticodon / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D012344:RNA, Transfer, Ala / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D000926:Anticodon; D012344:RNA, Transfer, Ala; D000412:Alanine-tRNA Ligase; D000409:Alanine