Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(R...Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(Reiter and Leroux,2017;Wallmeier et al.,2020).To date,over 180 ciliopathy-associated genes have been identified(Reiter and Leroux,2017),yet the underlying mechanisms remain poorly understood.展开更多
The study of the neuron has always been a fundamental aspect when it came to studying mental illnesses such as autism and depression. The protein protocadherin-9 (PCDH9) is an important transmembrane protein in the de...The study of the neuron has always been a fundamental aspect when it came to studying mental illnesses such as autism and depression. The protein protocadherin-9 (PCDH9) is an important transmembrane protein in the development of the neuron synapse. Hence, research on its protein interactome is key to understanding its functionality and specific properties. A newly discovered biotin ligase, TurboID, is a proximity labeler that is designed to be able to label and observe transmembrane proteins, something that previous methods struggled with. The TurboID method is verified in HEK293T cells and primary cultured mouse cortical neurons. Results have proven the validity of the TurboID method in observing PCDH9-interacting proteins.展开更多
Germ granules are biomolecular condensates composed of RNA and proteins that play crucial roles in RNA metabolism and post-transcriptional gene regulation.C.elegans germ granules consist of multiple distinct subcompar...Germ granules are biomolecular condensates composed of RNA and proteins that play crucial roles in RNA metabolism and post-transcriptional gene regulation.C.elegans germ granules consist of multiple distinct subcompartments,including P granules,Mutator foci,Z granules,SIMR foci,P-bodies,D granules,and E granules.Among these condensates,the E granule,which is nonrandomly positioned within the germ granule,is required for the production of a specialized class of small interfering RNAs(siRNAs).However,the mechanisms underlying E granule formation and its functional significance remain largely unexplored.In this study,via the use of TurboID-based proximity labeling technology combined with an RNAi-based reverse genetic screen,we identified two novel components of the E granule,EGC-2/C27B7.5 and EGC-3/F59G1.8,which initiate E granule assembly.The depletion of EGC-2 or EGC-3 disrupts the perinuclear localization of the EGO and PICS complexes,both of which are enriched in E granules and are required for E-class siRNA and piRNA biogenesis,respectively.Small RNAomic analyses revealed that both EGC-2 and EGC-3 promote the production of 5′E-class siRNA,whereas piRNA accumulation is inhibited by EGC-3.Taken together,our results elucidate the roles of EGC-2 and EGC-3 in maintaining E granule integrity and small RNA homeostasis.Additionally,the combination of proximity labeling technology and reverse genetic screening provides a robust strategy for studying the assembly of biomolecular condensates.展开更多
基金supported by grants from the National Key Research and Development Program of China(2019YFA0802704)the National Natural Science Foundation of China(31771620)+2 种基金the Natural Science Foundation of Chongqing,China(CSTB2022NSCQMSX1424)Research Startup Fund of Southwest University(SWU117064)Open Research Fund of National Health Commission Key Laboratory of Birth Defects Prevention&Henan Key Laboratory of Population Defects Prevention(ZD202302)。
文摘Cilia are indispensable for organ development and function,and their dysfunction causes a range of syndromic diseases known as ciliopathies,including obesity,cystic kidney disease,situs inversus,and male infertility(Reiter and Leroux,2017;Wallmeier et al.,2020).To date,over 180 ciliopathy-associated genes have been identified(Reiter and Leroux,2017),yet the underlying mechanisms remain poorly understood.
文摘The study of the neuron has always been a fundamental aspect when it came to studying mental illnesses such as autism and depression. The protein protocadherin-9 (PCDH9) is an important transmembrane protein in the development of the neuron synapse. Hence, research on its protein interactome is key to understanding its functionality and specific properties. A newly discovered biotin ligase, TurboID, is a proximity labeler that is designed to be able to label and observe transmembrane proteins, something that previous methods struggled with. The TurboID method is verified in HEK293T cells and primary cultured mouse cortical neurons. Results have proven the validity of the TurboID method in observing PCDH9-interacting proteins.
基金supported by the National Key Research and Development Program of China(2022YFA1302700)the National Natural Science Foundation of China(32230016,32270583,32470633,32400435,2023M733425,32300438)+2 种基金the Research Funds of Center for Advanced Interdisciplinary Science and Biomedicine of IHM(QYPY20230021)the Fundamental Research Funds for the Central UniversitiesSome strains were provided by the CGC,which is funded by the NIH Office of Research Infrastructure Programs(P40 OD010440).
文摘Germ granules are biomolecular condensates composed of RNA and proteins that play crucial roles in RNA metabolism and post-transcriptional gene regulation.C.elegans germ granules consist of multiple distinct subcompartments,including P granules,Mutator foci,Z granules,SIMR foci,P-bodies,D granules,and E granules.Among these condensates,the E granule,which is nonrandomly positioned within the germ granule,is required for the production of a specialized class of small interfering RNAs(siRNAs).However,the mechanisms underlying E granule formation and its functional significance remain largely unexplored.In this study,via the use of TurboID-based proximity labeling technology combined with an RNAi-based reverse genetic screen,we identified two novel components of the E granule,EGC-2/C27B7.5 and EGC-3/F59G1.8,which initiate E granule assembly.The depletion of EGC-2 or EGC-3 disrupts the perinuclear localization of the EGO and PICS complexes,both of which are enriched in E granules and are required for E-class siRNA and piRNA biogenesis,respectively.Small RNAomic analyses revealed that both EGC-2 and EGC-3 promote the production of 5′E-class siRNA,whereas piRNA accumulation is inhibited by EGC-3.Taken together,our results elucidate the roles of EGC-2 and EGC-3 in maintaining E granule integrity and small RNA homeostasis.Additionally,the combination of proximity labeling technology and reverse genetic screening provides a robust strategy for studying the assembly of biomolecular condensates.
