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Neural Regeneration Research 2023年第9期18卷 1952-1953页

作者： Stuart J.Grice ji-long liu Medical Research Council Functional Genomics Unit Department of PhysiologyAnatomy and GeneticsUniversity of OxfordOxfordUK School of Life Science and Technology ShanghaiTech UniversityShanghaiChina

Developmental motifs in neurodegeneration:Neurodegeneration,the prominent feature of neurodegenerative disease,is characterized by the progressive and selective loss of neuronal function.As some of the pathologies caused by neurodegeneration may be irreversible,early intervention will be required for the treatments that aim to slow or halt the manifestation of these diseases.Traditionally,neurodegeneration evokes the idea of a progressive decline of brain function.

关键词： neurodegeneration Huntington function

Temperature-sensitive cytoophidium assembly in Schizosaccharomyces pombe

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Journal of Genetics and Genomics 2019年第9期46卷 423-432页

作者： jing Zhang ji-long liu MRC Functional Genomics Unit Department of PhysiologyAnatomy and GeneticsUniversity of OxfordOxford0X13PTUnited Kingdom School of Life Science and Technology ShanghaiTech UniversityShanghai201210China

The metabolic enzyme CTP synthase(CTPS) is able to compartmentalize into filaments,termed cytoophidia,in a variety of organisms including bacteria,budding yeast,fission yeast,fruit flies and mammals.A previous study in budding yeast shows that the filament-forming process of CTPS is not sensitive to temperature shift.Here we study CTPS filamentation in the fission yeast Schizosaccharomyces pombe.To our surprise,we find that both the length and the occurrence of cytoophidia in S.pombe decrease upon cold shock or heat shock.The temperature-dependent changes of cytoophidia are fast and reversible.Taking advantage of yeast genetics,we demonstrate that heat-shock proteins are required for cytoophidium assembly in S.pombe.Temperature sensitivity of cytoophidia makes S.pombe an attractive model system for future investigations of this novel membraneless organelle.

关键词： CTP synthase Cytoophidium Schizosaccharomyces pombe Heat-shock protein Nucleoside/nucleotide metabolism Cell biology Yeast genetics Cell compartmentalization

mTOR-S6K1 pathway mediates cytoophidium assembly

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Journal of Genetics and Genomics 2019年第2期46卷 65-74页

作者： Zhe Sun ji-long liu School of Life Science and Technology ShanghaiTech University MRC Functional Genomics Unit Department of Physiology Anatomy and Genetics University of Oxford

CTP synthase(CTPS), the rate-limiting enzyme in de novo CTP biosynthesis, has been demonstrated to assemble into evolutionarily conserved filamentous structures, termed cytoophidia, in Drosophila, bacteria, yeast and mammalian cells. However, the regulation and function of the cytoophidium remain elusive. Here, we provide evidence that the mechanistic target of rapamycin(mTOR) pathway controls cytoophidium assembly in mammalian and Drosophila cells. In mammalian cells, we find that inhibition of mTOR pathway attenuates cytoophidium formation. Moreover, CTPS cytoophidium assembly appears to be dependent on the mTOR complex 1(mTORC1) mainly. In addition, knockdown of the mTORC1 downstream target S6 K1 can inhibit cytoophidium formation, while overexpression of the constitutively active S6 K1 reverses mTOR knockdown-induced cytoophidium disassembly. Finally, reducing m TOR protein expression results in a decrease of the length of cytoophidium in Drosophila follicle cells.Therefore, our study connects CTPS cytoophidium formation with the mTOR signaling pathway.

关键词： mTOR Cytoophidium CTP synthase Colorectal cancer cell Drosophila

The atlas of cytoophidia in Drosophila larvae

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Journal of Genetics and Genomics 2020年第6期47卷 321-331页

作者： Yuanbing Zhang jingnan liu ji-long liu School of Life Science and Technology Shanghai Tech UniversityShanghai201210China University of Chinese Academy of Sciences Beijing100049China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai200031China

In 2010,cytidine 50-triphosphate synthase(CTPS)was reported to form the filamentous or serpentine structure in Drosophila,which we termed the cytoophidium.In the last decade,CTPS filaments/cytoophidia have been found in bacteria,budding yeast,human cells,mice,fission yeast,plants,and archaea,indicating that this mechanism is highly conserved in evolution.In addition to CTPS,other metabolic enzymes have been identified to have the characteristics of forming cytoophidia or similar advanced structures,demonstrating that this is a basic strategy of cells.Nevertheless,our understanding of the physiological function of the cytoophidium remains incomplete and elusive.Here,we took the larva of Drosophila melanogaster as a model to systematically describe the localization and distribution of cytoophidia in different tissues during larval development.We found that the distribution pattern of CTPS cytoophidia is dynamic and heterogenic in larval tissues.Our study provides a road map for further understanding of the function and regulatory mechanism of cytoophidia.

关键词： CTP synthase Cytoophidium Drosophila Larva Metabolic enzyme

CTP synthase forms cytoophidia in archaea

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Journal of Genetics and Genomics 2020年第4期47卷 213-223页

作者： Shuang Zhou Hua Xiang ji-long liu School of Life Science and Technology ShanghaiTech UniversityShanghai201210China University of Chinese Academy of Sciences Beijing100049China Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai200031China State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing100101China

CTP synthase(CTPS)is an important metabolic enzyme that catalyzes the rate-Iimiting reaction of nucleotide CrP de novo synthesis.Since 2010,a series of studies have demonstrated that CTPS can form filamentous structures in bacteria and eukaryotes,which are termed cytoophidia.However,it is unknown whether cytoophidia exist in the third domain of life,archaea.Using Haloarcula hispanica as a model system,here we demonstrate that CTPS forms distinct intracellular compartments in archaea.Under stimulated emission depletion microscopy,we find that the structures of H.hispanica CTPS are elongated,similar to cytoophidia in bacteria and eukaryotes.When Haloarcula cells are cultured in lowsalt medium,the occurrence of cytoophidia increases dramatically.In addition,treatment of H.hispanica with a glutamine analog or overexpression of CTPS can promote cytoophidium assembly.Our study reveals that CTPS can fo rm cytoophidia in all three domains of life,suggesting that forming cytoophidia is an ancient property of CTPS.

关键词： Archaea CTP synthase Cytoophidium Haloarcula hispanica

Sparks of the CRISPR explosion:Applications in medicine and agriculture

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Journal of Genetics and Genomics 2017年第9期44卷 413-414页

作者： ji-long liu School of Life Science and Technology ShanghaiTech University Department of Physiology Anatomy and Genetics University of Oxford

Exactly 30 years ago,Nakata and coworkers published a paper analyzing the 1664-nucleotide sequence of a chromosomal DNA segment that contained the iap gene and its flanking regions in Escherichia coli（Ishino et al.,1... 详细信息

关键词： Sparks of the CRISPR explosion:Applications in medicine and agriculture

May the Force Be with You: Metabolism of Arginine and Pyrimidines

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Journal of Genetics and Genomics 2015年第5期42卷 179-180页

作者： ji-long liu Medical Research Council Functional Genomics Unit Department of Physiology Anatomy and Genetics University of Oxford

＂Why do arginine and pyrimidines have to be considered together？＂ This was the question I asked when I was invited by Barbara Zimmermann to attend the 23rd Intemational Conference on Arginine and Pyrimidines （ICAP） three years ago. The meeting was held in the summer of 2012 at Bogota, Columbia, known as the ＂the Athens of South America＂. I am not alone in wanting to know more about the relationship between arginine and pyrimidines. Last summer, when ! was organising the next ICAP meeting at Oxford （Aughey et al., 2014）, one of my colleagues asked me, ＂Why such a weird name for a conference？＂ I am not sure if I had given her a satisfying answer and I was enthused to find out what the reasons may be.

关键词： ICAP Metabolism of Arginine and Pyrimidines May the Force Be with You CPS

CRISPR/Cas9 and Genome Editing in Drosophila

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Journal of Genetics and Genomics 2014年第1期41卷 7-19页

作者： Andrew R.Bassett ji-long liu MRC Functional Genomics Unit University of Oxford Department of Physiology Anatomy and Genetics

Recent advances in our ability to design DNA binding factors with specificity for desired sequences have resulted in a revolution in genetic engineering, enabling directed changes to the genome to be made relatively easily. Traditional techniques for generating genetic mutations in most organisms have relied on selection from large pools of randomly induced mutations for those of particular interest, or time-consuming gene targeting by homologous recombination. Drosophila melanogaster has always been at the forefront of genetic analysis, and application of these new genome editing techniques to this organism will revolutionise our approach to performing analysis of gene function in the future. We discuss the recent techniques that apply the CRISPR/Cas9 system to Drosophila, highlight potential uses for this technology and speculate upon the future of genome engineering in this model organism.

关键词： Drosophila melanogaster CRISPR Cas9 Genome engineering Targeted mutagenesis

CTP Synthase Is Required for Optic Lobe Homeostasis in Drosophila

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Journal of Genetics and Genomics 2015年第5期42卷 261-274页

作者： ?mür Y.Tastan ji-long liu Medical Research Council Functional Genomics Unit Department of PhysiologyAnatomy and GeneticsUniversity of Oxford

CTP synthase （CTPsyn） is a metabolic enzyme responsible for the de novo synthesis of the nucleotide CTE Several recent studies have shown that CTPsyn forms filamentous subcellular structures known as cytoophidia in bacteria, yeast, fruit flies and humans. However, it remains elusive whether and how CTPsyn and cytoophidia play a role during development. Here, we show that cytoophidia are abundant in the neuroepithelial stem cells in Drosophila optic lobes. Optic lobes are underdeveloped in CTPsyn mutants as well as in CTPsyn RNAi. Moreover, overexpressing CTPsyn impairs the development of optic lobes, specifically by blocking the transition from neuro- epithelium to neuroblast. Taken together, our results indicate that CTPsyn is critical for optic lobe homeostasis in Drosophila.

关键词： CTP synthase Cytoophidium Drosophila Central nervous system Neuroepithelial stem cell

A Genome-Wide CRISPR Library for High-Throughput Genetic Screening in Drosophila Cells

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Journal of Genetics and Genomics 2015年第6期42卷 301-309页

作者： Andrew R.Bassett Lesheng Kong ji-long liu MRC Functional Genomics Unit University of OxfordDepartment of PhysiologyAnatomy and Genetics Genome Engineering Oxford Sir William Dunn School of PathologyUniversity of Oxford

The simplicity of the CRISPR/Cas9 system of genome engineering has opened up the possibility of performing genome-wide targeted mutagenesis in cell lines,enabling screening for cellular phenotypes resulting from genetic aberrations.Drosophila cells have proven to be highly effective in identifying genes involved in cellular processes through similar screens using partial knockdown by RNAi.This is in part due to the lower degree of redundancy between genes in this organism,whilst still maintaining highly conserved gene networks and orthologs of many human disease-causing genes.The ability of CRISPR to generate genetic loss of function mutations not only increases the magnitude of any effect over currently employed RNAi techniques,but allows analysis over longer periods of time which can be critical for certain phenotypes.In this study,we have designed and built a genome-wide CRISPR library covering 13,501 genes,among which 8989 genes are targeted by three or more independent single guide RNAs（sg RNAs）.Moreover,we describe strategies to monitor the population of guide RNAs by high throughput sequencing（HTS）.We hope that this library will provide an invaluable resource for the community to screen loss of function mutations for cellular phenotypes,and as a source of guide RNA designs for future studies.

关键词： CRISPR/Cas9 Genome-wide library Drosophila