Mapping contacts between Escherichia coli alanyl tRNA synthetase and 2 ' hydroxyls using a complete tRNA molecule

Citation
Ja. Pleiss et al., Mapping contacts between Escherichia coli alanyl tRNA synthetase and 2 ' hydroxyls using a complete tRNA molecule, BIOCHEM, 39(28), 2000, pp. 8250-8258
Citations number
74
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMISTRY
ISSN journal
00062960 → ACNP
Volume
39
Issue
28
Year of publication
2000
Pages
8250 - 8258
Database
ISI
SICI code
0006-2960(20000718)39:28<8250:MCBECA>2.0.ZU;2-M
Abstract
A dual-specific derivative of yeast tRNA(Phe) is described whose features f acilitate structure-function studies of tRNAs. This tRNA has been made in t hree different bimolecular forms that allow modifications to be easily intr oduced into any position within the molecule. A set of deoxynucleotide subs tituted versions of this tRNA has been created and used to examine contacts between tRNA and Escherichia coli alanyl-tRNA synthetase, an enzyme previo usly shown to interact with 2'-hydroxyls in the acceptor stem of the tRNA. Because the present experiments used a full-length tRNA, several contacts w ere identified that had not been previously found using microhelix substrat es. Contacts at similar sites in the T-loop are seen in the cocrystal struc ture of tRNA(Ser) and Thermus thermophilus seryl-tRNA synthetase.