Title

Sustained Translational Repression of Lactate Dehydrogenase 1 in Toxoplasma gondii Bradyzoites is Conferred by a Small Regulatory RNA Hairpin.

Document Type

Article

Publication Date

11-2014

Publication Title

FEBS J

Volume

281

Issue

22

First Page

5077

Last Page

5091

DOI

10.1111/febs.13048

Keywords

50UTR, lactate dehydrogenase, RNA hairpin, Toxoplasma gondii, translational repression

Abstract

In response to environmental stresses, Toxoplasma gondii induces a global translational repression which allows for the remodeling of its transcriptome. While some transcripts are preferentially translated, another subset is translationally repressed and maintained in bradyzoites. Although little is known of how transcripts are targeted for sustained translational repression, the targeting probably operates through an RNA-centric mechanism relying on the recognition of cis-acting elements. In this study, we sought to determine if the targeting of transcripts through recognizable cis-acting elements could be responsible for the transcript-specific sustained translational repression displayed by Toxoplasma bradyzoites. We examined the UTRs of a translationally repressed gene, lactate dehydrogenase 1, and found a 40 nucleotide regulatory element in its 5'UTR. This element specifically induces translational repression in otherwise constitutively expressed transcripts. Mutational studies revealed that the formation of a small 16 nucleotide regulatory RNA hairpin is essential for this activity. We suggest that this hairpin may act as the nucleation site for the binding of an as yet to be identified trans-acting factor that allows for the transcript to be targeted for translational repression removal from the active translational pool. To our knowledge, this is the first report characterizing a specific cis-acting element contributing to post-transcriptional gene regulation in Toxoplasma and suggests the presence of a pathway by which the parasites can recognize, identify and specifically target transcripts for sustained translational repression under stressful conditions.

Funding Reference Number

222969

Comments

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) to SA (grant number 222969). MH was supported by a graduate student fellowship from the Ministry of Col- leges and Universities, Ontario, Canada. The authors would like to thank Vaso Globarevic for technical assistance as well as Anna K. Crater and Emad Manni for critical analysis.