DOI: 10.1073/pnas.85.4.1033. Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.

Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.

10.1073/pnas.85.4.1033

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

A recombinant plasmid was constructed with six synthetic DNA oligomers such that the DNA sequence corresponding to yeast tRNA(Phe) is flanked by a T7 promoter and a BstNI restriction site. Runoff transcription of the BstNI-digested plasmid with T7 RNA polymerase gives an unmodified tRNA of the expected sequence having correct 5' and 3' termini. This tRNA(Phe) transcript can be specifically aminoacylated by yeast phenylalanyl-tRNA synthetase and has a Km only 4-fold higher than that of the native yeast tRNA(Phe). The Km is independent of Mg2+ concentration, whereas the Vmax is very dependent on Mg2+ concentration. Comparison of the melting profiles of the native and the unmodified tRNA(Phe) at different Mg2+ concentrations suggests that the unmodified tRNA(Phe) has a less stable tertiary structure. Using one additional DNA oligomer, a mutant plasmid was constructed having a guanosine to thymidine change at position 20 in the tRNA gene. A decrease in Vmax/Km by a factor of 14 for aminoacylation of the mutant tRNA(Phe) transcript is observed.

Authors 2

J R Sampson (first)
O C Uhlenbeck (additional)

References 0 Referenced 390

None

Dates

Type	When
Created	19 years, 2 months ago (May 31, 2006, 6:49 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 12:37 p.m.)
Indexed	1 month ago (July 16, 2025, 9:25 a.m.)
Issued	37 years, 6 months ago (Feb. 1, 1988)
Published	37 years, 6 months ago (Feb. 1, 1988)
Published Online	37 years, 6 months ago (Feb. 1, 1988)
Published Print	37 years, 6 months ago (Feb. 1, 1988)

Funders 0

None

BibTeX

@article{Sampson_1988, title={Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.}, volume={85}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.85.4.1033}, DOI={10.1073/pnas.85.4.1033}, number={4}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Sampson, J R and Uhlenbeck, O C}, year={1988}, month=feb, pages={1033–1037} }

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