Conserved requirement for DEAD-box RNA helicase Gemin3 in Drosophila oogenesis

Cauchi, Ruben J.

doi:10.1186/1756-0500-5-120

Cited by 11 publications

(11 citation statements)

References 50 publications

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“…Consistent with the well-documented role of RNA helicases in oogenesis and fertility (87,(123)(124)(125)(126), we identified two putative DEAD-box helicases, CG10077 and Mahe (Maheshvara). While CG10077 is an ortholog of human DDX5, Mahe is an evolutionary conserved regulator of Notch signaling (127).…”

Section: Identification Of Novel Genes With a Potential Role In Drososupporting

confidence: 83%

An interaction network of RNA-binding proteins involved inDrosophilaoogenesis

Bansal

Madlung

Schaaf

et al. 2020

Preprint

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bioRxiv preprint Interactome of RNA Binding Proteins from Drosophila ovaries 2 Abbreviations used EGFP enhanced green fluorescent protein eIF eukaryotic initiation factor EJC exon junction complex FDR false discovery rate GO gene ontology HA Hemagglutinin hnRNP heterogeneous nuclear ribonucleoproteins IP immunoprecipitation LFQ label-free quantification MBP maltose-binding protein mRNP messenger ribonucleoprotein NMD nonsense-mediated decay PFA paraformaldehyde piRNA piwi-interacting RNA RBP RNA-binding proteins SILAC stable isotope labeling with amino acids UAS upstream activating sequence AbstractDuring Drosophila oogenesis, the localization and translational regulation of maternal transcripts relies on RNA-binding proteins (RBPs). Many of these RBPs localize several mRNAs and may have additional direct interaction partners to regulate their functions. Using immunoprecipitation from whole Drosophila ovaries coupled to mass spectrometry, we examined protein-protein associations of 6 GFP-tagged RBPs expressed at physiological levels. Analysis of the interaction network and further validation in human cells allowed us to identify 26 previously unknown associations, besides recovering several well characterized interactions. We identified interactions between RBPs and several splicing factors, providing links between nuclear and cytoplasmic events of mRNA regulation. Additionally, components of the translational and RNA decay machineries were selectively co-purified with some baits, suggesting a mechanism for how RBPs may regulate maternal transcripts. Given the evolutionary conservation of the studied RBPs, the interaction network presented here provides the foundation for future functional and structural studies of mRNA localization across metazoans. : bioRxiv preprint Interactome of RNA Binding Proteins from Drosophila ovaries 5 consistent with the role of their mammalian homologs (38, 50-52). During early oogenesis, both Vas and Nos are involved in the maintenance of germline stem cells, in oocyte differentiation and other aspects of oocyte development (26, 35, 53-56). In embryos, Nos functions in germline development (53, 57-61) and further promotes the inclusion of germline cells in the developing ovary (53, 62). In addition to oogenic processes, Nos and Stau are also involved in the development of the Drosophila nervous system (63, 64). Many RBPs in Drosophila oogenesis have overlapping functions that are likely differentially regulated. Little is known about this regulation and it may involve several as yet unidentified mRNP components. To comprehensively identify RBP interactors, we carried out a systematic in vivo purification screen of GFPtagged RBPs coupled with mass spectrometry. We employed both labeled and label-free MS methods and identified several proteins significantly enriched with the purified RBPs. The interactomes of the individual RBPs were largely independent with some overlap. Our screen identified several previously unknown interactions, many of which we validated in vitro. This work pres...

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Section: Identification Of Novel Genes With a Potential Role In Drososupporting

confidence: 83%

An interaction network of RNA-binding proteins involved inDrosophilaoogenesis

Bansal

Madlung

Schaaf

et al. 2020

Preprint

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“…In contrast to other DEAD box proteins that have been directly implicated in Drosophila oogenesis (Cauchi, 2012;Johnstone et al, 2005;Styhler et al, 1998;Tomancak et al, 1998), we propose that the infertility observed in Ddx1 null females is due to disrupted metabolism. Altered metabolism also provides a possible explanation for the small number of progeny generated by Ddx1 AX /Df (3L)ED230 females, as we observed slightly higher pS6k levels and slightly larger body size in these mutants compared to the completely sterile Ddx1 AX/AX females.…”

Section: Discussioncontrasting

confidence: 63%

Loss of the Drosophila melanogaster DEAD box protein Ddx1 leads to reduced size and aberrant gametogenesis

Germain

Hildebrandt

et al. 2015

Developmental Biology

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Mammalian DDX1 has been implicated in RNA trafficking, DNA double-strand break repair and RNA processing; however, little is known about its role during animal development. Here, we report phenotypes associated with a null Ddx1 (Ddx1(AX)) mutation generated in Drosophila melanogaster. Ddx1 null flies are viable but significantly smaller than control and Ddx1 heterozygous flies. Female Ddx1 null flies have reduced fertility with egg chambers undergoing autophagy, whereas males are sterile due to disrupted spermatogenesis. Comparative RNA sequencing of control and Ddx1 null third instars identified several transcripts affected by Ddx1 inactivation. One of these, Sirup mRNA, was previously shown to be overexpressed under starvation conditions and implicated in mitochondrial function. We demonstrate that Sirup is a direct binding target of Ddx1 and that Sirup mRNA is differentially spliced in the presence or absence of Ddx1. Combining Ddx1 null mutation with Sirup dsRNA-mediated knock-down causes epistatic lethality not observed in either single mutant. Our data suggest a role for Drosophila Ddx1 in stress-induced regulation of splicing.

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“…In addition, defects in the U2-snRNP component noisette produce a range of oogenic defects depending on the severity of the allele, including novel abnormal condensation of NC nuclei, delayed development, sensory bristle growth, and fertility [57]. Mutations in SMN also produce NCCD defects in the female germ line, which also suggest that impaired snRNP assembly may be a factor in the NCCD phenotype; the RNA-helicase Gemin 3 also co-localizes with SMN and retains NCCD failure in the mutant germ line as well [58], [59]. Therefore, defects in spliceosome integrity/dynamics can have dramatic effects on multiple aspects of development.…”

Section: Discussionmentioning

confidence: 99%

Prp22 and Spliceosome Components Regulate Chromatin Dynamics in Germ-Line Polyploid Cells

et al. 2013

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During Drosophila oogenesis, the endopolyploid nuclei of germ-line nurse cells undergo a dramatic shift in morphology as oogenesis progresses; the easily-visible chromosomes are initially polytenic during the early stages of oogenesis before they transiently condense into a distinct ‘5-blob’ configuration, with subsequent dispersal into a diffuse state. Mutations in many genes, with diverse cellular functions, can affect the ability of nurse cells to fully decondense their chromatin, resulting in a ‘5-blob arrest’ phenotype that is maintained throughout the later stages of oogenesis. However, the mechanisms and significance of nurse-cell (NC) chromatin dispersal remain poorly understood. Here, we report that a screen for modifiers of the 5-blob phenotype in the germ line isolated the spliceosomal gene peanuts, the Drosophila Prp22. We demonstrate that reduction of spliceosomal activity through loss of peanuts promotes decondensation defects in NC nuclei during mid-oogenesis. We also show that the Prp38 spliceosomal protein accumulates in the nucleoplasm of nurse cells with impaired peanuts function, suggesting that spliceosomal recycling is impaired. Finally, we reveal that loss of additional spliceosomal proteins impairs the full decondensation of NC chromatin during later stages of oogenesis, suggesting that individual spliceosomal subcomplexes modulate expression of the distinct subset of genes that are required for correct morphology in endopolyploid nurse cells.

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Conserved requirement for DEAD-box RNA helicase Gemin3 in Drosophila oogenesis

Cited by 11 publications

References 50 publications

An interaction network of RNA-binding proteins involved inDrosophilaoogenesis

An interaction network of RNA-binding proteins involved inDrosophilaoogenesis

Loss of the Drosophila melanogaster DEAD box protein Ddx1 leads to reduced size and aberrant gametogenesis

Prp22 and Spliceosome Components Regulate Chromatin Dynamics in Germ-Line Polyploid Cells

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