Enzymatic and structural characterization of rTSγ provides insights into the function of rTSβ

Daniel J. Wichelecki, D. Sean Froese, Jolanta Kopec, Joao R.C. Muniz, Wyatt W. Yue, John Alan Gerlt

Research output: Contribution to journalArticle

Abstract

In humans, the gene encoding a reverse thymidylate synthase (rTS) is transcribed in the reverse direction of the gene encoding thymidylate synthase (TS) that is involved in DNA biosynthesis. Three isoforms are found: α, β, and γ, with the transcript of the α-isoform overlapping with that of TS. rTSβ has been of interest since the discovery of its overexpression in methotrexate and 5-fluorouracil resistant cell lines. Despite more than 20 years of study, none of the rTS isoforms have been biochemically or structurally characterized. In this study, we identified rTSγ as an l-fuconate dehydratase and determined its high-resolution crystal structure. Our data provide an explanation for the observed difference in enzymatic activities between rTSβ and rTSγ, enabling more informed proposals for the possible function of rTSβ in chemotherapeutic resistance.

Original languageEnglish (US)
Pages (from-to)2732-2738
Number of pages7
JournalBiochemistry
Volume53
Issue number16
DOIs
StatePublished - Apr 29 2014

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Thymidylate Synthase
Protein Isoforms
Gene encoding
Hydro-Lyases
Biosynthesis
Methotrexate
Fluorouracil
Genes
Crystal structure
Cells
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wichelecki, D. J., Froese, D. S., Kopec, J., Muniz, J. R. C., Yue, W. W., & Gerlt, J. A. (2014). Enzymatic and structural characterization of rTSγ provides insights into the function of rTSβ. Biochemistry, 53(16), 2732-2738. https://doi.org/10.1021/bi500349e

Enzymatic and structural characterization of rTSγ provides insights into the function of rTSβ. / Wichelecki, Daniel J.; Froese, D. Sean; Kopec, Jolanta; Muniz, Joao R.C.; Yue, Wyatt W.; Gerlt, John Alan.

In: Biochemistry, Vol. 53, No. 16, 29.04.2014, p. 2732-2738.

Research output: Contribution to journalArticle

Wichelecki, DJ, Froese, DS, Kopec, J, Muniz, JRC, Yue, WW & Gerlt, JA 2014, 'Enzymatic and structural characterization of rTSγ provides insights into the function of rTSβ', Biochemistry, vol. 53, no. 16, pp. 2732-2738. https://doi.org/10.1021/bi500349e
Wichelecki, Daniel J. ; Froese, D. Sean ; Kopec, Jolanta ; Muniz, Joao R.C. ; Yue, Wyatt W. ; Gerlt, John Alan. / Enzymatic and structural characterization of rTSγ provides insights into the function of rTSβ. In: Biochemistry. 2014 ; Vol. 53, No. 16. pp. 2732-2738.
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