Modulation of Aub–TDRD interactions elucidates piRNA amplification and germplasm formation

Vrettos, Nicholas; Maragkakis, Manolis; Alexiou, Panagiotis; Sgourdou, Paraskevi; Ibrahim, Fadia; Palmieri, Daniel E.; Kirino, Yohei; Mourelatos, Zissimos P.

doi:10.26508/lsa.202000912

Cited by 10 publications

(14 citation statements)

References 53 publications

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“…Germ granules assemble at the posterior of D. melanogaster oocytes and their mRNAs are essential for primordial germ cell specification in the developing embryos (57). By genetic deletion or germ cell specific knockdown of Drosophila Prmt5 , we found that germ granules never formed as Aub with unmethylated NTRs could not interact with Tudor (51). piRNA biogenesis, amplification and transposon control were intact in flies expressing endogenous Aub with unmethylated NTRs (51).…”

Section: Discussionmentioning

confidence: 98%

“…By genetic deletion or germ cell specific knockdown of Drosophila Prmt5 , we found that germ granules never formed as Aub with unmethylated NTRs could not interact with Tudor (51). piRNA biogenesis, amplification and transposon control were intact in flies expressing endogenous Aub with unmethylated NTRs (51). However, by mutating Aub-NTRs to lysines, we found that germ granule formation, piRNA amplification and transposon control all collapsed, as the Aub-RK mutant protein was unable to interact not only with Tudor but also with Tdrds involved in piRNA amplification (51).…”

Section: Discussionmentioning

confidence: 98%

“…Our findings draw parallels between the functions of mouse MIWI/TDRDs and D. melanogaster Aub/Tdrds. We recently examined, genetically, the role of Aub-NTRs and their methylation status in piRNA biogenesis, amplification and formation of germ granules (51). The latter consist of mRNPs that contain mRNAs, bound directly by Aub (50), and also contain Tudor (50) (52) (53), Vasa (54) (55) (56) and other RNA-binding Proteins (57).…”

Section: Discussionmentioning

confidence: 99%

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MIWI arginines orchestrate generation of functional pachytene piRNAs and spermiogenesis

Vrettos,

Oppelt,

Zoch

et al. 2024

Preprint

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N-terminal arginine (NTR) methylation is a conserved feature of PIWI proteins, which are central components of the PIWI-interacting RNA (piRNA) pathway. The significance and precise function of PIWI NTR methylation in mammals remains unknown. In mice, PIWI NTRs bind Tudor domain containing proteins (TDRDs) that have essential roles in piRNA biogenesis and the formation of the chromatoid body. Using mouse MIWI (PIWIL1) as paradigm, we demonstrate that the NTRs are essential for spermatogenesis through the regulation of transposons and gene expression. Surprisingly, the loss of TDRD5 and TDRKH interaction with MIWI results in defective piRNA amplification, rather than an expected failure of piRNA biogenesis. We find that piRNA amplification is necessary for both transposon control and for sustaining levels of select, nonconserved, pachytene piRNAs that target specific mRNAs required for spermatogenesis. Our findings support the notion that the vast majority of pachytene piRNAs are dispensable, acting as autonomous genetic elements that rely for propagation on MIWI piRNA amplification. MIWI-NTRs also mediate interactions with TDRD6 that are necessary for chromatoid body compaction. Furthermore, MIWI-NTRs promote stabilization of spermiogenic transcripts that drive nuclear compaction, which is essential for sperm formation. In summary, the NTRs underpin the diversification of MIWI protein function.

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

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MIWI arginines orchestrate generation of functional pachytene piRNAs and spermiogenesis

Vrettos,

Oppelt,

Zoch

et al. 2024

Preprint

Self Cite

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“…Phasing of CDS-piRNAs collected from two libraries (Aub-IP and GFP-Aub-IP in cyp40 KO/6 background) was analyzed by the published protocol (74). Analyzed sRNA data were summarized in table S1 (75)(76)(77)(78)(79).…”

Section: Deep-seq Of Srnas and Data Processingmentioning

confidence: 99%

miRNA/siRNA-directed pathway to produce noncoding piRNAs from endogenous protein-coding regions ensures Drosophila spermatogenesis

2023

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PIWI-interacting RNA (piRNA) pathways control transposable elements (TEs) and endogenous genes, playing important roles in animal gamete formation. However, the underlying piRNA biogenesis mechanisms remain elusive. Here, we show that endogenous protein coding sequences (CDSs), which are normally used for translation, serve as origins of noncoding piRNA biogenesis in Drosophila melanogaster testes. The product, namely, CDS-piRNAs, formed silencing complexes with Aubergine (Aub) in germ cells. Proximity proteome and functional analyses show that CDS-piRNAs and cluster/TE-piRNAs are distinct species occupying Aub, the former loading selectively relies on chaperone Cyclophilin 40. Moreover, Argonaute 2 (Ago2) and Dicer-2 activities were found critical for CDS-piRNA production. We provide evidence that Ago2-bound short interfering RNAs (siRNAs) and microRNAs (miRNAs) specify precursors to be processed into piRNAs. We further demonstrate that Aub is crucial in spermatid differentiation, regulating chromatins through mRNA cleavage. Collectively, our data illustrate a unique strategy used by male germ line, expanding piRNA repertoire for silencing of endogenous genes during spermatogenesis.

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“…In different animals, Tud domain-containing proteins interact with symmetrically dimethylated arginines (sDMAs) of Piwi proteins in germ granules (Kirino et al, 2009;Vagin et al, 2009). In particular, in Drosophila germline, methylated Aub associates with Tud domain proteins Tud and Krimper (Nishida et al, 2009;Kirino et al, 2010;Huang et al, 2021;Vrettos et al, 2021). In glial granules, both Piwi and Ago3 were identified; however, Aub protein was not detected in these granules.…”

Section: Piwi Family Proteinsmentioning

confidence: 99%

Looking at the Pretty “Phase” of Membraneless Organelles: A View From Drosophila Glia

Arkov

2022

Front. Cell Dev. Biol.

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Membraneless granules assemble in different cell types and cellular loci and are the focus of intense research due to their fundamental importance for cellular organization. These dynamic organelles are commonly assembled from RNA and protein components and exhibit soft matter characteristics of molecular condensates currently characterized with biophysical approaches and super-resolution microscopy imaging. In addition, research on the molecular mechanisms of the RNA–protein granules assembly provided insights into the formation of abnormal granules and molecular aggregates, which takes place during many neurodegenerative disorders including Parkinson’s diseases (PD), Alzheimer’s disease (AD), amyotrophic lateral sclerosis (ALS), and frontotemporal dementia (FTD). While these disorders are associated with formation of abnormal granules, membraneless organelles are normally assembled in neurons and contribute to translational control and affect stability of neuronal RNAs. More recently, a new subtype of membraneless granules was identified in Drosophila glia (glial granules). Interestingly, glial granules were found to contain proteins which are the principal components of the membraneless granules in germ cells (germ granules), indicating some similarity in the functional assembly of these structures in glia and germline. This mini review highlights recent research on glial granules in the context of other membraneless organelles, including their assembly mechanisms and potential functions in the nervous system.

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Modulation of Aub–TDRD interactions elucidates piRNA amplification and germplasm formation

Cited by 10 publications

References 53 publications

MIWI arginines orchestrate generation of functional pachytene piRNAs and spermiogenesis

MIWI arginines orchestrate generation of functional pachytene piRNAs and spermiogenesis

miRNA/siRNA-directed pathway to produce noncoding piRNAs from endogenous protein-coding regions ensures Drosophila spermatogenesis

Looking at the Pretty “Phase” of Membraneless Organelles: A View From Drosophila Glia

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