Musashi interaction with poly(A)-binding protein is required for activation of target mRNA translation

Chad E Cragle,Melanie C Macnicol,Stephanie D Byrum,Linda L Hardy,Samuel G Mackintosh,William A Richardson,Nicola K Gray,Gwen V Childs,Alan J Tackett,Angus M Macnicol

doi:10.1074/jbc.ra119.007220

Abstract

The Musashi family of mRNA translational regulators controls both physiological and pathological stem cell self-renewal primarily by repressing target mRNAs that promote differentiation. In response to differentiation cues, Musashi can switch from a repressor to an activator of target mRNA translation. However, the molecular events that distinguish Musashi-mediated translational activation from repression are not understood. We have previously reported that Musashi function is required for the maturation of Xenopus oocytes and specifically for translational activation of specific dormant maternal mRNAs. Here, we employed MS to identify cellular factors necessary for Musashi-dependent mRNA translational activation. We report that Musashi1 needs to associate with the embryonic poly(A)-binding protein (ePABP) or the canonical somatic cell poly(A)-binding protein PABPC1 for activation of Musashi target mRNA translation. Co-immunoprecipitation studies demonstrated an increased Musashi1 interaction with ePABP during oocyte maturation. Attenuation of endogenous ePABP activity severely compromised Musashi function, preventing downstream signaling and blocking oocyte maturation. Ectopic expression of either ePABP or PABPC1 restored Musashi-dependent mRNA translational activation and maturation of ePABP-attenuated oocytes. Consistent with these Xenopus findings, PABPC1 remained associated with Musashi under conditions of Musashi target mRNA de-repression and translation during mammalian stem cell differentiation. Because association of Musashi1 with poly(A)-binding proteins has previously been implicated only in repression of Musashi target mRNAs, our findings reveal novel context-dependent roles for the interaction of Musashi with poly(A)-binding protein family members in response to extracellular cues that control cell fate.

Highlights

The Musashi family of mRNA translational regulators controls both physiological and pathological stem cell self-renewal primarily by repressing target mRNAs that promote differentiation
We demonstrate that Musashi1 interacts with several members of the poly(A)-binding protein family, including embryonic poly(A)-binding protein (ePABP) and PABPC1, to promote translational activation of target mRNAs
Our mapping data indicate that Musashi1 possesses two domains for interaction with ePABP, whereas in reciprocal experiments Musashi1 was shown to interact with the C-terminal domain of both ePABP and PABPC1

Summary

Introduction

The Musashi family of mRNA translational regulators controls both physiological and pathological stem cell self-renewal primarily by repressing target mRNAs that promote differentiation. Ectopic expression of either ePABP or PABPC1 restored Musashi-dependent mRNA translational activation and maturation of ePABP-attenuated oocytes. Consistent with these Xenopus findings, PABPC1 remained associated with Musashi under condi-. Because association of Musashi with poly(A)-binding proteins has previously been implicated only in repression of Musashi target mRNAs, our findings reveal novel context-dependent roles for the interaction of Musashi with poly(A)-binding protein family members in response to extracellular cues that control cell fate. As a consequence of this competition, it was suggested that the Musashi1–PABPC1 interaction prevented recruitment of the large 60S ribosomal subunit and subsequent cap-dependent translation of Musashi target mRNAs [27]

Methods

Results

Conclusion