1887

Abstract

Thymidylate synthase (TS) enzymes catalyse the biosynthesis of deoxythymidine monophosphate (dTMP or thymidylate), and so are important for DNA replication and repair. Two different types of TS proteins have been described (ThyA and ThyX), which have different enzymic mechanisms and unrelated structures. Mycobacteria are unusual as they encode both and , and the biological significance of this is not yet understood. ThyX is thought to be essential and a potential drug target. We therefore analysed and expression levels, their essentiality and roles in pathogenesis. We show that both and are expressed , and that this expression significantly increased within murine macrophages. Under all conditions tested, expression exceeded that of . Mutational studies show that is essential, confirming that the enzyme is a plausible drug target. The requirement for in the presence of implies that the essential function of ThyX is something other than dTMP synthase. We successfully deleted from the genome, and this deletion conferred an growth defect that was not observed . Presumably ThyX performs TS activity within Δ at a sufficient rate for normal growth, but the rate is less than optimal. We also demonstrate that deletion confers -aminosalicylic acid resistance, and show by complementation studies that ThyA T202A and V261G appear to be functional and non-functional, respectively.

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2012-02-01
2019-12-05
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