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JOURNAL ARTICLE
Xiaowei Guo, Yihao Sun, Taha Azad, H J Janse van Rensburg, Jingjing Luo, Shuai Yang, Peng Liu, Zhongwei Lv, Meixiao Zhan, Ligong Lu, Yingqun Zhou, Xianjue Ma, Xiaoping Zhang, Xiaolong Yang, Lei Xue
The Hippo pathway is an evolutionarily conserved regulator of organ growth and tumorigenesis. In Drosophila , oncogenic RasV12 cooperates with loss-of-cell polarity to promote Hippo pathway-dependent tumor growth. To identify additional factors that modulate this signaling, we performed a genetic screen utilizing the Drosophila Ras V12 /lgl -/- in vivo tumor model and identified Rox8, a RNA-binding protein (RBP), as a positive regulator of the Hippo pathway. We found that Rox8 overexpression suppresses whereas Rox8 depletion potentiates Hippo-dependent tissue overgrowth, accompanied by altered Yki protein level and target gene expression...
November 17, 2020: Proceedings of the National Academy of Sciences of the United States of America
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JOURNAL ARTICLE
Sebastian Wurster, Alexander M Tatara, Nathaniel D Albert, Ashraf S Ibrahim, Joseph Heitman, Soo Chan Lee, Amol C Shetty, Carrie McCracken, Karen T Graf, Antonios G Mikos, Vincent M Bruno, Dimitrios P Kontoyiannis
Trauma-related necrotizing myocutaneous mucormycosis (NMM) has a high morbidity and mortality in victims of combat-related injuries, geometeorological disasters, and severe burns. Inspired by the observation that several recent clusters of NMM have been associated with extreme mechanical forces (e.g., during tornados), we studied the impact of mechanical stress on Mucoralean biology and virulence in a Drosophila melanogaster infection model. In contrast to other experimental procedures to exert mechanical stress, tornadic shear challenge (TSC) by magnetic stirring induced a hypervirulent phenotype in several clinically relevant Mucorales species but not in Aspergillus or Fusarium Whereas fungal growth rates, morphogenesis, and susceptibility to noxious environments or phagocytes were not altered by TSC, soluble factors released in the supernatant of shear-challenged R...
June 30, 2020: MBio
#23
JOURNAL ARTICLE
Prashali Bansal, Johannes Madlung, Kristina Schaaf, Boris Macek, Fulvia Bono
During 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 Drosophilaovaries 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...
June 17, 2020: Molecular & Cellular Proteomics: MCP
#24
JOURNAL ARTICLE
Lisenka E L M Vissers, Sreehari Kalvakuri, Elke de Boer, Sinje Geuer, Machteld Oud, Inge van Outersterp, Michael Kwint, Melde Witmond, Simone Kersten, Daniel L Polla, Dilys Weijers, Amber Begtrup, Kirsty McWalter, Anna Ruiz, Elisabeth Gabau, Jenny E V Morton, Christopher Griffith, Karin Weiss, Candace Gamble, James Bartley, Hilary J Vernon, Kendra Brunet, Claudia Ruivenkamp, Sarina G Kant, Paul Kruszka, Austin Larson, Alexandra Afenjar, Thierry Billette de Villemeur, Kimberly Nugent, F Lucy Raymond, Hanka Venselaar, Florence Demurger, Claudia Soler-Alfonso, Dong Li, Elizabeth Bhoj, Ian Hayes, Nina Powell Hamilton, Ayesha Ahmad, Rachel Fisher, Myrthe van den Born, Marjolaine Willems, Arthur Sorlin, Julian Delanne, Sebastien Moutton, Philippe Christophe, Frederic Tran Mau-Them, Antonio Vitobello, Himanshu Goel, Lauren Massingham, Chanika Phornphutkul, Jennifer Schwab, Boris Keren, Perrine Charles, Maaike Vreeburg, Lenika De Simone, George Hoganson, Maria Iascone, Donatella Milani, Lucie Evenepoel, Nicole Revencu, D Isum Ward, Kaitlyn Burns, Ian Krantz, Sarah E Raible, Jill R Murrell, Kathleen Wood, Megan T Cho, Hans van Bokhoven, Maximilian Muenke, Tjitske Kleefstra, Rolf Bodmer, Arjan P M de Brouwer
CNOT1 is a member of the CCR4-NOT complex, which is a master regulator, orchestrating gene expression, RNA deadenylation, and protein ubiquitination. We report on 39 individuals with heterozygous de novo CNOT1 variants, including missense, splice site, and nonsense variants, who present with a clinical spectrum of intellectual disability, motor delay, speech delay, seizures, hypotonia, and behavioral problems. To link CNOT1 dysfunction to the neurodevelopmental phenotype observed, we generated variant-specific Drosophila models, which showed learning and memory defects upon CNOT1 knockdown...
July 2, 2020: American Journal of Human Genetics
#25
JOURNAL ARTICLE
Keon Mook Seong, YeongHo Kim, Donghun Kim, Barry R Pittendrigh, Young Ho Kim
The fruit fly, Drosophila melanogaster, is predominantly found in overripe, rotten, fermenting, or decaying fruits and is constantly exposed to chemical stressors such as acetic acid, ethanol, and 2-phenylethanol. D. melanogaster has been employed as a model system for studying the molecular bases of various types of chemical-induced tolerance. Expression profiling using Illumina sequencing has been performed for identifying changes in gene expression that may be associated with evolutionary adaptation to exposure of acetic acid, ethanol, and 2-phenylethanol...
May 2020: Pesticide Biochemistry and Physiology
#26
JOURNAL ARTICLE
André Hallen, Arthur J L Cooper
A novel cosegregating splice site variant in the Dynactin-1 ( DCTN1 ) gene was discovered by Next Generation Sequencing (NGS) in a family with a history of bipolar disorder (BD) and major depressive diagnosis (MDD). Psychiatric illness in this family follows an autosomal dominant pattern. DCTN1 codes for the largest dynactin subunit, namely p150Glued , which plays an essential role in retrograde axonal transport and in neuronal autophagy. A GT→TT transversion in the DCTN1 gene, uncovered in the present work, is predicted to disrupt the invariant canonical splice donor site IVS22 + 1G > T and result in intron retention and a premature termination codon (PTC)...
April 18, 2020: Genes
#27
JOURNAL ARTICLE
René M Arvola, Chung-Te Chang, Joseph P Buytendorp, Yevgen Levdansky, Eugene Valkov, Peter L Freddolino, Aaron C Goldstrohm
Pumilio is an RNA-binding protein that represses a network of mRNAs to control embryogenesis, stem cell fate, fertility and neurological functions in Drosophila. We sought to identify the mechanism of Pumilio-mediated repression and find that it accelerates degradation of target mRNAs, mediated by three N-terminal Repression Domains (RDs), which are unique to Pumilio orthologs. We show that the repressive activities of the Pumilio RDs depend on specific subunits of the Ccr4-Not (CNOT) deadenylase complex. Depletion of Pop2, Not1, Not2, or Not3 subunits alleviates Pumilio RD-mediated repression of protein expression and mRNA decay, whereas depletion of other CNOT components had little or no effect...
December 21, 2019: Nucleic Acids Research
#28
JOURNAL ARTICLE
Burak Gür, Katja Sporar, Anne Lopez-Behling, Marion Silies
The computational organization of sensory systems depends on the diversification of individual cell types with distinct signal-processing capabilities. The Drosophila visual system, for instance, splits information into channels with different temporal properties directly downstream of photoreceptors in the first-order interneurons of the OFF pathway, L2 and L3. However, the biophysical mechanisms that determine this specialization are largely unknown. Here, we show that the voltage-gated Ka channels Shaker and Shal contribute to the response properties of the major OFF pathway input L2...
December 10, 2019: Journal of Comparative Physiology. A, Neuroethology, Sensory, Neural, and Behavioral Physiology
#29
JOURNAL ARTICLE
Mandy Li-Ian Tay, Jun Wei Pek
Gene expression involves the transcription and splicing of nascent transcripts through the removal of introns. In Drosophila, a double-stranded RNA binding protein Disco-interacting protein 1 (DIP1) targets INE-1 stable intronic sequence RNAs (sisRNAs) for degradation after splicing. How nascent transcripts that also contain INE-1 sequences escape degradation remains unknown. Here we observe that these nascent transcripts can also be bound by DIP1 but the Drosophila homolog of SON (Dsn) protects them from unproductive degradation in ovaries...
November 15, 2019: PLoS Genetics
#30
JOURNAL ARTICLE
Aoife Hanet, Felix Räsch, Ramona Weber, Vincenzo Ruscica, Maria Fauser, Tobias Raisch, Duygu Kuzuoğlu-Öztürk, Chung-Te Chang, Dipankar Bhandari, Cátia Igreja, Lara Wohlbold
Eukaryotic superfamily (SF) 1 helicases have been implicated in various aspects of RNA metabolism, including transcription, processing, translation, and degradation. Nevertheless, until now, most human SF1 helicases remain poorly understood. Here, we have functionally and biochemically characterized the role of a putative SF1 helicase termed "helicase with zinc-finger," or HELZ. We discovered that HELZ associates with various mRNA decay factors, including components of the carbon catabolite repressor 4-negative on TATA box (CCR4-NOT) deadenylase complex in human and Drosophila melanogaster cells...
October 2019: Life Science Alliance
#31
JOURNAL ARTICLE
Brian Reichholf, Veronika A Herzog, Nina Fasching, Raphael A Manzenreither, Ivica Sowemimo, Stefan L Ameres
Argonaute-bound microRNAs silence mRNA expression in a dynamic and regulated manner to control organismal development, physiology, and disease. We employed metabolic small RNA sequencing for a comprehensive view on intracellular microRNA kinetics in Drosophila. Based on absolute rate of biogenesis and decay, microRNAs rank among the fastest produced and longest-lived cellular transcripts, disposing up to 105 copies per cell at steady-state. Mature microRNAs are produced within minutes, revealing tight intracellular coupling of biogenesis that is selectively disrupted by pre-miRNA-uridylation...
August 22, 2019: Molecular Cell
#32
JOURNAL ARTICLE
Vincenzo Ruscica, Praveen Bawankar, Daniel Peter, Sigrun Helms, Cátia Igreja, Elisa Izaurralde
The eIF4E-homologous protein (4EHP) is a translational repressor that competes with eIF4E for binding to the 5'-cap structure of specific mRNAs, to which it is recruited by protein factors such as the GRB10-interacting GYF (glycine-tyrosine-phenylalanine domain) proteins (GIGYF). Several experimental evidences suggest that GIGYF proteins are not merely facilitating 4EHP recruitment to transcripts but are actually required for the repressor activity of the complex. However, the underlying molecular mechanism is unknown...
May 22, 2019: Nucleic Acids Research
#33
JOURNAL ARTICLE
Wangchao Xu, Puhua Bao, Xin Jiang, Haifang Wang, Meiling Qin, Ruiqi Wang, Tao Wang, Yi Yang, Ileana Lorenzini, Lujian Liao, Rita Sattler, Jin Xu
Amyotrophic lateral sclerosis is a deleterious neurodegenerative disease without effective treatment options. Recent studies have indicated the involvement of the dysregulation of RNA metabolism in the pathogenesis of amyotrophic lateral sclerosis. Among the various RNA regulatory machineries, nonsense-mediated mRNA decay (NMD) is a stress responsive cellular surveillance system that degrades selected mRNA substrates to prevent the translation of defective or harmful proteins. Whether this pathway is affected in neurodegenerative diseases is unclear...
April 1, 2019: Brain
#34
JOURNAL ARTICLE
Anand K Singh, Subhendu Roy Choudhury, Sandip De, Jie Zhang, Stephen Kissane, Vibha Dwivedi, Preethi Ramanathan, Marija Petric, Luisa Orsini, Daniel Hebenstreit, Saverio Brogna
UPF1 is an RNA helicase that is required for nonsense-mediated mRNA decay (NMD) in eukaryotes, and the predominant view is that UPF1 mainly operates on the 3'UTRs of mRNAs that are directed for NMD in the cytoplasm. Here we offer evidence, obtained from Drosophila , that UPF1 constantly moves between the nucleus and cytoplasm by a mechanism that requires its RNA helicase activity. UPF1 is associated, genome-wide, with nascent RNAs at most of the active Pol II transcription sites and at some Pol III-transcribed genes, as demonstrated microscopically on the polytene chromosomes of salivary glands and by ChIP-seq analysis in S2 cells...
March 25, 2019: ELife
#35
JOURNAL ARTICLE
Kazuko Hanyu-Nakamura, Kazuki Matsuda, Stephen M Cohen, Akira Nakamura
Specification of germ cells is pivotal to ensure continuation of animal species. In many animal embryos, germ cell specification depends on maternally supplied determinants in the germ plasm. Drosophila polar granule component ( pgc ) mRNA is a component of the germ plasm. pgc encodes a small protein that is transiently expressed in newly formed pole cells, the germline progenitors, where it globally represses mRNA transcription. pgc is also required for pole cell survival, but the mechanism linking transcriptional repression to pole cell survival remains elusive...
April 4, 2019: Development
#36
JOURNAL ARTICLE
Hwajung Ri, Jongbin Lee, Jun Young Sonn, Eunseok Yoo, Chunghun Lim, Joonho Choe
Post-transcriptional regulation underlies the circadian control of gene expression and animal behaviors. However, the role of mRNA surveillance via the nonsense-mediated mRNA decay (NMD) pathway in circadian rhythms remains elusive. Here, we report that Drosophila NMD pathway acts in a subset of circadian pacemaker neurons to maintain robust 24h rhythms of free-running locomotor activity. RNA interference-mediated depletion of key NMD factors in timeless-expressing clock cells decreased the amplitude of circadian locomotor behaviors...
March 28, 2019: Molecules and Cells
#37
JOURNAL ARTICLE
Zhaohui Li, Fu Yang, Yang Xuan, Rongwen Xi, Rui Zhao
Hbs1, which is homologous to the GTPase eRF3, is a small G protein implicated in mRNA quality control. It interacts with a translation-release factor 1-like protein Dom34/Pelota to direct decay of mRNAs with ribosomal stalls. Although both proteins are evolutionarily conserved in eukaryotes, the biological function of Hbs1 in multicellular organisms is yet to be characterized. In Drosophila, pelota is essential for the progression through meiosis during spermatogenesis and germline stem cell maintenance. Here we show that homozygous Hbs1 mutant flies are viable, female-fertile, but male-sterile, which is due to defects in meiosis and spermatid individualization, phenotypes that are also observed in pelota hypomorphic mutants...
March 1, 2019: Scientific Reports
#38
REVIEW
Zheng Xing, Wai Kit Ma, Elizabeth J Tran
The mammalian DEAD-box RNA helicase DDX5, its paralog DDX17, and their orthologs in Saccharomyces cerevisiae and Drosophila melanogaster, namely Dbp2 and Rm62, define a subfamily of DEAD-box proteins. Members from this subfamily share highly conserved protein sequences and cellular functions. They are involved in multiple steps of RNA metabolism including mRNA processing, microRNA processing, ribosome biogenesis, RNA decay, and regulation of long noncoding RNA activities. The DDX5/Dbp2 subfamily is also implicated in transcription regulation, cellular signaling pathways, and energy metabolism...
December 2, 2018: Wiley Interdisciplinary Reviews. RNA
#39
JOURNAL ARTICLE
Hideyuki Komori, Krista L Golden, Taeko Kobayashi, Ryoichiro Kageyama, Cheng-Yu Lee
Self-renewal genes maintain stem cells in an undifferentiated state by preventing the commitment to differentiate. Robust inactivation of self-renewal gene activity following asymmetric stem cell division allows uncommitted stem cell progeny to exit from an undifferentiated state and initiate the commitment to differentiate . Nonetheless, how self-renewal gene activity at mRNA and protein levels becomes synchronously terminated in uncommitted stem cell progeny is unclear. We demonstrate that a multilayered gene regulation system terminates self-renewal gene activity at all levels in uncommitted stem cell progeny in the fly neural stem cell lineage...
November 21, 2018: Genes & Development
#40
JOURNAL ARTICLE
Evelina Tutucci, Maria Vera, Robert H Singer
The MS2 system has been widely used, in organisms ranging from bacteria to higher eukaryotes, to image single mRNAs in intact cells with high precision. We have recently re-engineered the MS2 system for accurate detection of mRNAs in living Saccharomyces cerevisiae. Previous MS2 systems affected the degradation of the tagged mRNA, which led to accumulation of MS2 fragments and to erroneous conclusions about mRNA localization and expression. Here we describe a step-by-step protocol for the use of our latest MS2 system (MBSV6) for detecting endogenously tagged mRNAs using wide-field fluorescent microscopy in living yeast...
October 2018: Nature Protocols
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