Binding interactions between yeast tRNA ligase and a precursor transfer ribonucleic acid containing two photoreactive uridine analogues

Journal Article · Tue Nov 29 00:00:00 EST 1988 · Biochemistry; (United States)

OSTI ID:6023841

Tanner, N K; Hanna, M M; Abelson, J

Yeast tRNA ligase, from Saccharomyces cerevisiae, is one of the protein components that is involved in the splicing reaction of intron-containing yeast precursor tRNAs. It is an unusual protein because it has three distinct catalytic activities. It functions as a polynucleotide kinase, as a cyclic phosphodiesterase, and as an RNA ligase. We have studied the binding interactions between ligase and precursor tRNAs containing two photoreactive uridine analogues, 4-thiouridine and 5-bromouridine. When irradiated with long ultraviolet light, RNA containing these analogues can form specific covalent bonds with associated proteins. In this paper, we show that 4-thiouridine triphosphate and 5-bromouridine triphosphate were readily incorporated into a precursor tRNA(Phe) that was synthesized, in vitro, with bacteriophage T7 RNA polymerase. The analogue-containing precursor tRNAs were authentic substrates for the two splicing enzymes that were tested (endonuclease and ligase), and they formed specific covalent bonds with ligase when they were irradiated with long-wavelength ultraviolet light. We have determined the position of three major cross-links and one minor cross-link on precursor tRNA(Phe) that were located within the intron and near the 3' splice site. On the basis of these data, we present a model for the in vivo splicing reaction of yeast precursor tRNAs.

• Research Organization: California Institute of Technology, Pasadena (USA)
• OSTI ID: 6023841
• Journal Information: Biochemistry; (United States), Vol. 27:24
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
LIGASES
BIOCHEMICAL REACTION KINETICS
TRANSFER RNA
CHEMICAL RADIATION EFFECTS
CROSS-LINKING
MATHEMATICAL MODELS
PHOSPHODIESTERASES
PHOSPHOTRANSFERASES
PRECURSOR
REAGENTS
RNA POLYMERASES
SACCHAROMYCES CEREVISIAE
ULTRAVIOLET RADIATION
URIDINE
AZINES
CHEMICAL REACTIONS
CHEMISTRY
ELECTROMAGNETIC RADIATION
ENZYMES
ESTERASES
EUMYCOTA
FUNGI
HETEROCYCLIC COMPOUNDS
HYDROLASES
HYDROXY COMPOUNDS
KINETICS
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PLANTS
POLYMERASES
POLYMERIZATION
PYRIMIDINES
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
REACTION KINETICS
RIBOSIDES
RNA
SACCHAROMYCES
TRANSFERASES
URACILS
YEASTS
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture