Abstract
BackgroundRNA regulation by RNA-binding proteins (RBPs) involve extremely complicated mechanisms. MOV10 and MOV10L1 are two homologous RNA helicases implicated in distinct intracellular pathways. MOV10L1 participates specifically in Piwi-interacting RNA (piRNA) biogenesis and protects mouse male fertility. In contrast, the functional complexity of MOV10 remains incompletely understood, and its role in the mammalian germline is unknown. Here, we report a study of the biological and molecular functions of the RNA helicase MOV10 in mammalian male germ cells.ResultsMOV10 is a nucleocytoplasmic protein mainly expressed in spermatogonia. Knockdown and transplantation experiments show that MOV10 deficiency has a negative effect on spermatogonial progenitor cells (SPCs), limiting proliferation and in vivo repopulation capacity. This effect is concurrent with a global disturbance of RNA homeostasis and downregulation of factors critical for SPC proliferation and/or self-renewal. Unexpectedly, microRNA (miRNA) biogenesis is impaired due partially to decrease of miRNA primary transcript levels and/or retention of miRNA via splicing control. Genome-wide analysis of RNA targetome reveals that MOV10 binds preferentially to mRNAs with long 3′-UTR and also interacts with various non-coding RNA species including those in the nucleus. Intriguingly, nuclear MOV10 associates with an array of splicing factors, particularly with SRSF1, and its intronic binding sites tend to reside in proximity to splice sites.ConclusionsThese data expand the landscape of MOV10 function and highlight a previously unidentified role initiated from the nucleus, suggesting that MOV10 is a versatile RBP involved in a broader RNA regulatory network.
Highlights
RNA regulation by RNA-binding proteins (RBPs) involve extremely complicated mechanisms
MILI and MVH are known to be present in perinuclear granules in the cytoplasm. d Wholemount immunostaining for MOV10 and PLZF in seminiferous tubules
Western blot analysis of MOV10 protein in subcellular fractions prepared from P10 testes confirmed the presence of considerable amounts of MOV10 protein in the nuclear fraction (Fig. 1c), which contained only minimal levels of MILI and MVH, well-defined Piwi-interacting RNA (piRNA) factors that mainly associate with cytoplasmic perinuclear granules called nuage [55]
Summary
RNA regulation by RNA-binding proteins (RBPs) involve extremely complicated mechanisms. We report a study of the biological and molecular functions of the RNA helicase MOV10 in mammalian male germ cells. Recent studies have shown that the transcriptome of the mammalian testis is exceptionally complex [1], due to the involvement of a vast variety of non-coding RNAs (ncRNAs) [2, 3]. A third major class of regulatory RNA species, lncRNAs, are defined as non-protein coding transcripts (> 200 nt in length). Mammalian male germ cells express thousands of unique lncRNAs in developmental-specific patterns [3, 16], and the regulatory function of a subset has been characterized [17,18,19,20]. LncRNAs are expressed at much lower levels than transcripts of protein-coding genes, limiting comprehensive detection by standard approaches [15, 22]
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