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Translation of TNFAIP2 is tightly controlled by upstream open reading frames

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Abstract

Translation is a highly regulated process, both at the global as well as on a transcript-specific level. Regulatory upstream open reading frames (uORFs) represent a mode to alter cap-dependent translation efficiency in a transcript-specific manner and are found in numerous mRNAs. In the majority of cases, uORFs inhibit the translation of their associated main ORFs. Consequently, their inactivation results in enhanced translation of the main ORF, a phenomenon best characterized in the context of the integrated stress response. In the present study, we identified potent translation-inhibitory uORFs in the transcript leader sequence (TLS) of tumor necrosis factor alpha induced protein 2 (TNFAIP2). The initial description of the uORFs was based on the observation that despite a massive induction of TNFAIP2 mRNA expression in response to interleukin 1β (IL1β), TNFAIP2 protein levels remained low in MCF7 cells. While we were able to characterize the uORFs with respect to their exact size and sequential requirements in this cellular context, only TPA stimulation partially overcame the translation-inhibitory activity of the TNFAIP2 uORFs. Characterization of TNFAIP2 translation in the context of monocyte-to-macrophage differentiation suggested that, while the uORFs efficiently block TNFAIP2 protein synthesis in monocytes, they are inactivated in mature macrophages, thus allowing for a massive increase in TNFAIP2 protein expression. In summary, we establish TNFAIP2 as a novel target of uORF-mediated translational regulation. Furthermore, our findings suggest that during macrophage differentiation a major uORF-dependent translational switch occurs.

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Abbreviations

CDS:

Coding sequence

CHX:

Cycloheximide

DMSO:

Dimethyl sulfoxide

IL1β:

Interleukin 1β

ISR:

Integrated stress response

MΦ:

Macrophages

MO:

Monocytes

RPF:

Ribosome-protected fragments

TLS:

Transcript leader sequence

TNFAIP2:

Tumor necrosis factor α induced protein 2

TPA:

12-O-tetradecanoylphorbol-13-acetate

uORF:

Upstream open reading frame

UTR:

Untranslated region

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Acknowledgements

We thank Kathi Zarnack for discussion of the bioinformatics analyses.

Funding

This work was supported by the Deutsche Forschungsgemeinschaft (SCHM2663/3 and GRK2336) and a fellowship by the Stiftung Polytechnische Gesellschaft Frankfurt (to AS).

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  1. Faculty of Medicine, Institute of Biochemistry I, Goethe-University Frankfurt, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany

    Anica Scholz, Peter Rappl, Nicola Böffinger, Ana Carolina Mota, Bernhard Brüne & Tobias Schmid

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Scholz, A., Rappl, P., Böffinger, N. et al. Translation of TNFAIP2 is tightly controlled by upstream open reading frames. Cell. Mol. Life Sci. 77, 2017–2027 (2020). https://doi.org/10.1007/s00018-019-03265-4

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