sup 1 H- sup 15 N NMR studies of Escherichia coli tRNA sup Phe from hisT mutants: A structural role for pseudouridine

Journal Article · Tue Apr 30 00:00:00 EDT 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00231a017· OSTI ID:5397996

Davis, D R; Poulter, C D ^[1]

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1. Univ. of Utah, Salt Lake City (USA)

Escherichia coli tRNA{sub U39}{sup Phe} was isolated from a specially constructed bacterial strain (DD1003/pRK3) carrying mutations in the hisT gene (the structural gene for tRNA pseudouridine synthase I) and in the pyrB gene (uracil auxotrophy). The pheU gene for tRNA{sup Phe} under control of the native tRNA promoter was on a multicopy plasmid and gave up to 40-fold overproduction of tRNA{sub U39}{sup Phe}. The double mutant permitted efficient incorporation of (3-{sup 15}N)uracil, resulting in >95% {sup 15}N enrichment of the uracil-derived bases. {sup 1}H and {sup 1}H-{sup 15}N NMR experiments were used to assign the low-field proton resonances to specific hydrogen-bonding interactions. {sup 1}H NMR assignments indicate that tRNA{sub U39}{sup Phe} has a structure similar to that of native tRNA{sup Phe} except in the anticodon region where replacement of pseudouridine ({Psi}) at position 39 with uridine (U) destabilizes hydrogen-bonding interactions at the base of the anticodon stem. The authors propose that U {yields} {Psi} modifications further stabilize interactions normally available to U by providing an additional locus for hydrogen bonding to the pyrimidine ring.

• OSTI ID: 5397996
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:17; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
AZINES
BACTERIA
BARYONS
CELL CONSTITUENTS
CHEMICAL SHIFT
CHROMATOGRAPHY
ELEMENTARY PARTICLES
ENZYMES
ESCHERICHIA COLI
FERMIONS
GENE MUTATIONS
HADRONS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPES
LIGASES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
MAGNETIC RESONANCE
MICROORGANISMS
MOLECULAR STRUCTURE
MUTATIONS
NITROGEN 15
NITROGEN ISOTOPES
NUCLEAR MAGNETIC RESONANCE
NUCLEI
NUCLEIC ACIDS
NUCLEONS
NUCLEOSIDES
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PLASMIDS
PROTONS
PYRIMIDINES
RESONANCE
RIBOSIDES
RNA
SEPARATION PROCESSES
STABLE ISOTOPES
TRANSFER RNA
URACILS
URIDINE