Abstract
Post-transcriptional regulation of gene expression plays an essential role during oocyte maturation. Here we report that Drosophila MARF1 (Meiosis Regulator And mRNA Stability Factor 1), which consists of one RNA-recognition motif and six tandem LOTUS domains with unknown molecular function, is essential for oocyte maturation. When tethered to a reporter mRNA, MARF1 post-transcriptionally silences reporter expression by shortening reporter mRNA poly-A tail length and thereby reducing reporter protein level. This activity is mediated by the MARF1 LOTUS domain, which binds the CCR4-NOT deadenylase complex. MARF1 binds cyclin A mRNA and shortens its poly-A tail to reduce Cyclin A protein level during oocyte maturation. This study identifies MARF1 as a regulator in oocyte maturation and defines the conserved LOTUS domain as a post-transcriptional effector domain that recruits CCR4-NOT deadenylase complex to shorten target mRNA poly-A tails and suppress their translation.
Highlights
Post-transcriptional regulation of gene expression plays an essential role during oocyte maturation
We report that Drosophila MARF1 (Meiosis Regulator And mRNA Stability Factor 1), which consists of one RNA-recognition motif and six tandem LOTUS domains with unknown molecular function, is essential for oocyte maturation
We found that 3xHA-MARF1 coimmunoprecipitated with CCR4-NOT deadenylase complex components (Not[1], Not[2], Not[3], and Caf1) in an RNAindependent manner (Fig. 9a). 3xHA-MARF1 did not coimmunoprecipitate with endogenous MARF1, DEAD-box helicase Vasa, DEAD-box helicase Belle, DEAD-box helicase Me31B, Oskar, GW182, Ge-1, Cyclin A, Cyclin B, Cyclin B3, cyclin-dependent kinase 1 (CDK1), or alpha-Tubulin, showing that the interaction between MARF1 and the CCR4-NOT deadenylase complex is specific (Supplementary Fig. 5A)
Summary
Post-transcriptional regulation of gene expression plays an essential role during oocyte maturation. We report that Drosophila MARF1 (Meiosis Regulator And mRNA Stability Factor 1), which consists of one RNA-recognition motif and six tandem LOTUS domains with unknown molecular function, is essential for oocyte maturation. When tethered to a reporter mRNA, MARF1 post-transcriptionally silences reporter expression by shortening reporter mRNA poly-A tail length and thereby reducing reporter protein level This activity is mediated by the MARF1 LOTUS domain, which binds the CCR4-NOT deadenylase complex. This study identifies MARF1 as a regulator in oocyte maturation and defines the conserved LOTUS domain as a post-transcriptional effector domain that recruits CCR4-NOT deadenylase complex to shorten target mRNA poly-A tails and suppress their translation. We uncovered the biological and molecular functions of Drosophila MARF1 and defined its conserved LOTUS domains as a post-transcriptional effector domain to recruit the CCR4-NOT deadenylase complex to shorten target mRNA poly-A tails and suppress translation of the mRNAs
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