Novel activity of a yeast ligase deletion polypeptide: Evidence for GTP-dependent tRNA splicing

Shawn Westaway, Heather G. Belford, Barbara L. Apostol, John Abelson, Chris L. Greer

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Yeast tRNA ligase possesses multiple activities which are required for the joining of tRNA halves during the tRNA splicing process: cyclic phosphodiesterase, kinase, adenylylate synthetase, and ligase. A deletion polypeptide of a dihydrofolate reductase-ligase fusion protein, designated DAC, was previously shown to join tRNA halves although ATP-dependent kinase activity was not measurable in the assay used. We describe here a characterization of the mechanism of joining used by DAC and the structure of the tRNA product. DAC produces a joined tRNA and a splice junction with a structure identical to that produced by DAKC, the full-length dihydrofolate reductase-ligase fusion. Furthermore, DAC can use GTP as the sole cofactor in the joining reaction, in contrast to DAKC, which can only complete splicing in the presence of ATP Both enzymes exhibit GTP-dependent kinase activity at 100-fold greater efficiency than with ATP. These results suggest that a potential function for the center domain of tRNA ligase (missing in DAC) is to provide structural integrity and aid in substrate interactions and specificity. They also support the hypothesis that ligase may prefer to use two different cofactors during tRNA splicing.

Original languageEnglish (US)
Pages (from-to)2435-2443
Number of pages9
JournalJournal of Biological Chemistry
Volume268
Issue number4
StatePublished - Feb 5 1993
Externally publishedYes

Fingerprint

Ligases
Transfer RNA
Guanosine Triphosphate
Yeast
Yeasts
Peptides
Joining
Tetrahydrofolate Dehydrogenase
Phosphotransferases
Adenosine Triphosphate
RNA Ligase (ATP)
Fusion reactions
Phosphoric Diester Hydrolases
Structural integrity
Substrate Specificity
Assays
Substrates
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Westaway, S., Belford, H. G., Apostol, B. L., Abelson, J., & Greer, C. L. (1993). Novel activity of a yeast ligase deletion polypeptide: Evidence for GTP-dependent tRNA splicing. Journal of Biological Chemistry, 268(4), 2435-2443.

Novel activity of a yeast ligase deletion polypeptide : Evidence for GTP-dependent tRNA splicing. / Westaway, Shawn; Belford, Heather G.; Apostol, Barbara L.; Abelson, John; Greer, Chris L.

In: Journal of Biological Chemistry, Vol. 268, No. 4, 05.02.1993, p. 2435-2443.

Research output: Contribution to journalArticle

Westaway, S, Belford, HG, Apostol, BL, Abelson, J & Greer, CL 1993, 'Novel activity of a yeast ligase deletion polypeptide: Evidence for GTP-dependent tRNA splicing', Journal of Biological Chemistry, vol. 268, no. 4, pp. 2435-2443.
Westaway, Shawn ; Belford, Heather G. ; Apostol, Barbara L. ; Abelson, John ; Greer, Chris L. / Novel activity of a yeast ligase deletion polypeptide : Evidence for GTP-dependent tRNA splicing. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 4. pp. 2435-2443.
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