Temporally Regulated Traffic of HuR and Its Associated ARE-Containing mRNAs from the Chromatoid Body to Polysomes during Mouse Spermatogenesis

Mai Nguyen Chi,Bernard Jégou,Frédéric Chalmel,Khalid S A Khabar,Nathalie Vanzo,Dominique Morello,Eric Agius,Thomas Preiss

doi:10.1371/journal.pone.0004900

Abstract

BackgroundIn mammals, a temporal disconnection between mRNA transcription and protein synthesis occurs during late steps of germ cell differentiation, in contrast to most somatic tissues where transcription and translation are closely linked. Indeed, during late stages of spermatogenesis, protein synthesis relies on the appropriate storage of translationally inactive mRNAs in transcriptionally silent spermatids. The factors and cellular compartments regulating mRNA storage and the timing of their translation are still poorly understood. The chromatoid body (CB), that shares components with the P. bodies found in somatic cells, has recently been proposed to be a site of mRNA processing. Here, we describe a new component of the CB, the RNA binding protein HuR, known in somatic cells to control the stability/translation of AU-rich containing mRNAs (ARE-mRNAs).Methodology/Principal FindingsUsing a combination of cell imagery and sucrose gradient fractionation, we show that HuR localization is highly dynamic during spermatid differentiation. First, in early round spermatids, HuR colocalizes with the Mouse Vasa Homolog, MVH, a marker of the CB. As spermatids differentiate, HuR exits the CB and concomitantly associates with polysomes. Using computational analyses, we identified two testis ARE-containing mRNAs, Brd2 and GCNF that are bound by HuR and MVH. We show that these target ARE-mRNAs follow HuR trafficking, accumulating successively in the CB, where they are translationally silent, and in polysomes during spermatid differentiation.Conclusions/SignificanceOur results reveal a temporal regulation of HuR trafficking together with its target mRNAs from the CB to polysomes as spermatids differentiate. They strongly suggest that through the transport of ARE-mRNAs from the CB to polysomes, HuR controls the appropriate timing of ARE-mRNA translation. HuR might represent a major post-transcriptional regulator, by promoting mRNA storage and then translation, during male germ cell differentiation.

Highlights

Spermatogenesis is a highly regulated process whereby the spermatogonial stem cells at the basal side of the seminiferous tubules divide and differentiate to give rise to spermatozoa
AU-rich element (ARE)-transcriptome is widely expressed in male germ cells Mammalian AREs are cis-elements located in the 39 untranslated region (39 UTR) of many mRNAs with a diverse functional repertoire [16], for which they ensure accurate transport, stability, storage and translation in somatic cells [17]
In contrast to what is observed in most somatic cells, a large quantity of mRNAs accumulated during transcription are stored as mRNPs, which are subsequently assembled into polysomes [31]

Summary

Introduction

Spermatogenesis is a highly regulated process whereby the spermatogonial stem cells at the basal side of the seminiferous tubules divide and differentiate to give rise to spermatozoa. In contrast to most somatic tissues, where transcription and translation are concomitant, mRNA transcription and protein synthesis are temporally disconnected in the male germ cells. Late-stage specific protein synthesis relies on the appropriate storage of translationally inactive mRNAs in transcriptionally silent germ cells [3]. Many encode RNA-binding proteins (RBPs) [4], arguing that regulated mRNA storage, stabilization and translation are needed to ensure stage-specific protein synthesis. A temporal disconnection between mRNA transcription and protein synthesis occurs during late steps of germ cell differentiation, in contrast to most somatic tissues where transcription and translation are closely linked. During late stages of spermatogenesis, protein synthesis relies on the appropriate storage of translationally inactive mRNAs in transcriptionally silent spermatids. We describe a new component of the CB, the RNA binding protein HuR, known in somatic cells to control the stability/translation of AU-rich containing mRNAs (ARE-mRNAs)

Methods

Results

Conclusion