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chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

Journal of Animal Science and Biotechnology 2024年第1期15卷 171-181页

作者： Estel Viñolas-Vergés Jordi Ribas-Maynou Isabel Barranco Camila Peres Rubio Sergi Bonet Jordi Roca Marc Yeste Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaGironaSpain Biotechnology of Animal and Human Reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGironaSpain Department of Medicine and Animal Surgery Faculty of Veterinary ScienceUniversity of MurciaMurciaSpain Catalan Institution for Research and Advanced Studies(ICREA) S08010 BarcelonaSpain

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo *** some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF ***,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been *** This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;Pchromatin condensation than the *** could be related to the OSi of each fraction.

关键词： chromatin Condensation DNA integrity Ejaculate fractions Pig Protamination Sperm

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chromatin and epigenetic regulation of the telomerase reverse transcriptase gene

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Protein & Cell 2010年第1期1卷 22-32页

作者： Jiyue Zhu Yuanjun Zhao Shuwen Wang Department of Cellular and Molecular Physiology Pennsylvania State University College of MedicineHersheyPA 17033USA

Telomerase expression and telomere maintenance are critical for long-term cell proliferation and survival,and they play important roles in development,aging,and *** evidence has indicated that regulation of the rate-limiting subunit of human telomerase reverse transcriptase gene(hTERT)is a complex process in normal cells and many cancer *** addition to a number of transcriptional activators and repressors,the chromatin environment and epigenetic status of the endogenous hTERT locus are also pivotal for its regulation in normal human somatic cells and in tumorigenesis.

关键词： telomerase transcriptional regulation chromatin epigenetic regulation cancer

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chromatin and Nucleosome Organizations and DNA Replication of Nucleus Reassembled in vitro Using Purified Exogenous DNA and Xenopus Egg Extracts

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Science China Chemistry 1994年第6期37卷 677-690页

作者：张传茂张博翟中和 Department of Biology Peking University Beijing 100871 PRC

It has been demonstrated in the last ten years that the nuclear reassembly may occur in the cell-free systems from frog egg extracts added with exogenous naked DNA. However, there remains an open question : is the cell-free reassembled nucleus structurally similar to the nucleus in the intact cell ? That is, does the cell-free reassembled nucleus contain nucleosomes and chromatin? For this issue, we have designed experiments for identifying the internal structures of the cell-free reassembled nucleus. These experiments show that the nucleus reassembled in vitro also contains chromatin which is composed of typical 10 nm nucleosome fibers of 'beads-on-a-string', 30 nm filaments and the next higher-order structures. The digestion experiment with the enzyme micrococcal nuclease has demonstrated that the DNA in the nucleosome of the reconstituted chromatin is about 200 base pairs (bp) in length, of which 165 bp may be in the nucleosome particle, and 35 bp may be in the linker between two particles. Prolonging digestion of the 165-bp particle DNA fragment will yield a 146-bp fragment, which may be wounded in the nucleosome core. Experiments also provide evidence that the chromatin reconstitution in the cell-free reassembled nucleus is a progressive process, and that the nucleus can replicate its DNA. Based on these observations, we suppose that the chromatin of the cell-free reassembled nucleus may be structurally and functionally similar to the chromatin of the intact cells.

关键词： cell-free system nuclear reassembly chromatin reconstitution nucleosome DNA replication.

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Induction of systemic lupus erythematosus syndrome in BALB/c mice by immunization with active chromatin

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Acta Pharmacologica Sinica 2004年第6期25卷 105-109页

作者： Hong LI, Yun-yi ZHANG, Ya-nan SUN, Xi-yi HUANG, Yong-feng JIA, Duan LI2 Department of Pharmacology, Schoolof Pharmacy, Fudan University, Shanghai 200032, China Department of Pharmacology School of Pharmacy Fudan University Shanghai 200032 China

AIM: To establish an animal model for systemic lupus erythematosus (SLE)-like syndrome in mice. METHODS: BALB/c mice were immunized with active chromatin isolated from ConA-actived syngeneic spleno-lymphocytes. Plasma samples of mice were tested by enzyme-linked immunosorbent assays (ELISA) for the presence of IgG anti-dsDNA, -ssDNA, and anti-histone antibodies. Tumor necrosis factor-α (TNF-α) in serum was measured by ELISA. Spleno-lymphocyte proliferation assays and the levels of interferon-γ (IFN-γ) in supernatants were tested respectively. Proteinuria was measured. Kidneys were examined by direct immunohistochemical method and light microscopy. RESULTS: Anti-ds DNA, ssDNA, and histone antibodies were induced in active chromatin-immu- nized mice, the proliferation response of splenocytes to ConA and LPS were reduced, levels of interferon-γ in supernatants and TNF-α in serum were lowered. Lupus nephritis was assessed by the presence of Ig deposits, glomerular pathology and proteinuria. CONCLUSION: The active chromatin-induced SLE-like mouse model was similar to idiopathic SLE in human.

关键词： systemic lupus erythematosus chromatin autoantibody tumor necrosis factor interferon type II B-lymphocytes T-lymphocytes female enzyme-linked immunosorbent assay

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Acidic domains differentially read histone H3 lysine 4 methylation status and are widely present in chromatin-associated proteins

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Science China(Life Sciences) 2017年第2期60卷 138-151页

作者： Meng Wu Wei Wei Jiwei Chen Rong Cong Tieliu Shi Jiwen Li Jiemin Wong James X.Du Shanghai Key Laboratory of Regulatory Biology the Institute of Biomedical Sciences and School of Life SciencesEast China Normal UniversityShanghai 200241China Collaborative Innovation Center for Cancer Medicine Sun Yat-Sen University Cancer CenterGuangzhou 510060China University de Lyon Ecole Normale Supdrieure de LyonCNRS USR 3010Laboratoire Joliot-CurieLyon 69364France

Histone methylation is believed to provide binding sites for specific reader proteins, which translate histone code into biological function. Here we show that a family of acidic domain-containing proteins including nucleophosmin （NPM 1）, pp32, SET/TAF 113, nucleolin （NCL） and upstream binding factor （UBF） are novel H3K4me2-binding proteins. These proteins exhibit a unique pattern of interaction with methylated H3K4, as their binding is stimulated by H3K4me2 and inhibited by H3K4mel and H3K4me3. These proteins contain one or more acidic domains consisting mainly of aspartic and/or glutamic residues that are necessary for preferential binding of H3K4me2. Furthermore, we demonstrate that the acidic domain with sufficient length alone is capable of binding H3K4me2 in vitro and in vivo. NPM1, NCL and UBF require their acidic domains for association with and transcriptional activation ofrDNA genes. Interestingly, by defining acidic domain as a sequence with at least 20 acidic residues in 50 continuous amino acids, we identified 655 acidic domain-containing protein coding genes in the human genome and Gene Ontology （GO） analysis showed that many of the acidic domain proteins have chromatin-related functions. Our data suggest that acidic domain is a novel histone binding motif that can differentially read the status of H3K4 methylation and is broadly present in chromatin-associated proteins.

关键词： histone methylation H3K4mel H3K4me2 H3K4me3 acidic domain histone code transcription chromatin

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Tissue-specific Hi-C analyses of rice, foxtail millet and maize suggest non-canonical function of plant chromatin domains

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Journal of Integrative Plant Biology 2020年第2期62卷 201-217页

作者： Pengfei Dong Xiaoyu Tu Haoxuan Li Jianhua Zhang Donald Grierson Pinghua Li Silin Zhong State Key Laboratory of Agrobiotechnology School of Life SciencesThe Chinese University of Hong KongHong KongChina Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology Zhejiang UniversityHangzhou 310058China State Key Laboratory of Crop Biology College of AgronomyShandong Agricultural UniversityTai’an 271018China

chromatins are not randomly packaged in the nucleus and their organization plays important roles in transcription regulation,which is best studied in the mammalian *** in situ Hi-C,we have compared the 3D chromatin architectures of rice mesophyll and endosperm,foxtail millet bundle sheath and mesophyll,and maize bundle sheath,mesophyll and endosperm *** found that their global A/B compartment partitions are stable across tissues,while local A/B compartment has tissue-specific dynamic associated with differential gene *** domains are largely stable across tissues,while new domain border formations are often associated with transcriptional activation in the *** inside plant domains are not conserved across species,and lack significant co-expression behavior unlike those in mammalian *** we only observed chromatin loops between gene islands in the large genomes,the maize loop gene pairs’syntenic orthologs have shorter physical distances in small genome monocots,suggesting that loops instead of domains might have conserved biological *** study showed that plants’chromatin features might not have conserved biological functions as the mammalian ones.

关键词： plant chromatin domains Tissue-specific Hi-C analyses rice

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METTL14 is a chromatin regulator independent of its RNA N^(6)-methyladenosine methyltransferase activity

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Protein & Cell 2023年第9期14卷 683-697页

作者： Xiaoyang Dou Lulu Huang Yu Xiao Chang Liu Yini Li Xinning Zhang Lishan Yu Ran Zhao Lei Yang Chuan Chen Xianbin Yu Boyang Gao Meijie Qi Yawei Gao Bin Shen Shuying Sun Chuan He Jun Liu Department of Chemistry and Institute for Biophysical Dynamics University of ChicagoChicagoIL 60637USA Howard Hughes Medical Institute ChicagoIL 60637USA State Key Laboratory of Protein and Plant Gene Research Peking-Tsinghua Center for Life SciencesSchool of Life SciencesPeking UniversityBeijing 100871China Department of Physiology and Brain Science Institute Johns Hopkins University School of MedicineBaltimoreMD 21205USA Institute for Regenerative Medicine Shanghai East HospitalShanghai Key Laboratory of Signaling and Disease ResearchFrontier Science Center for Stem Cell ResearchSchool of Life Sciences and TechnologyTongji UniversityShanghai 200120China The Institute of Translational Medicine School of MedicineZhejiang UniversityHangzhou 310029China Division of Life Sciences and Medicine Center for Reproductive MedicineThe First Affiliated Hospital of USTCUniversity of Science and Technology of ChinaHefei230000China Clinical and Translational Research Center of Shanghai First Maternity&Infant Hospital Shanghai Key Laboratory of Signaling and Disease ResearchFrontier Science Center for Stem Cell ResearchSchool of Life Sciences and TechnologyTongji UniversityShanghai 200092China State Key Laboratory of Reproductive Medicine Center for Global HealthGusu SchoolWomen's Hospital of Nanjing Medical UniversityNanjing Maternity and Child Health Care HospitalNanjing Medical UniversityNanjing211166China Department of Biochemistry and Molecular Biology University of ChicagoChicagoIL 60637USA

METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m^(6)A methyltransferase complex(MTC)that installs m^(6)***,depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell(mESC).While comparable global hypo-methylation in RNA m^(6)A was observed in Mettl3 or Mettl14 knockout mESCs,***14 knockout led to a globally decreased nascent RNA synthesis,whereas Mettl3 depletion resulted in transcription upregulation,suggesting that METTL14 might possess an mA-independent role in gene *** found that METTL14 colocalizes with the repressive H3K27me3 ***,METTL14,but not METTL3,binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of *** of METTL14 thus led to a global increase in H3K27me3 level along with a global gene *** effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of *** work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m^(6)A,and critically impacts transcriptional regulation,stemness maintenance,and differentiation ofmESCs.

关键词： METTL14 chromatin H3K27me3 m^(6)A-independent mESC differentiation

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PCGF6 regulates stem cell pluripotency as a transcription activator via super-enhancer dependent chromatin interactions

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Protein & Cell 2019年第10期10卷 709-725页

作者： Xiaona Huang Chao Wei Fenjie Li Lumeng Jia Pengguihang Zeng Jiahe Li Jin Tan Tuanfeng Sun Shaoshuai Jiang Jia Wang Xiuxiao Tang Bin Liu Limin Rong Cheng Li Junjun Ding RNA Biomedical Institute Sun Yat-Sen Memorial HospitalZhongshan School of MedicineSun Yat-Sen UniversityGuangzhou 510080China Center for Stem Cell Biology and Tissue Engineering Key Laboratory for Stem Cells and Tissue EngineeringMinistry of EducationSun Yat-Sen UniversityGuangzhou 510080China Department of Cell Biology Zhongshan School of MedicineSun Yat-sen UniversityGuangzhou 510080China Program in Stem Cell and Regenerative Medicine The Third Affiliated Hospital of Sun Yat-sen UniversityZhongshan School of MedicineSun Yat-Sen UniversityGuangzhouChina Center for Bioinformatics School of Life SciencesPeking UniversityBeijing 100871China Department of Spine Surgery The Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhou 510630China Department of Histology and Embryology School of Basic Medical SciencesGuangzhou Medical UniversityGuangzhou 511436China

Polycomb group(PcG)ring finger protein 6(PCGF6),though known as a member of the transcription-re-pressing complexes,PcG,also has activation function in regulating pluripotency gene ***,the mechanism underlying the activation function of PCGF6 is poorly ***,we found that PCGF6 co-localizes to gene activation regions along with pluripotency factors such as *** addition,PCGF6 was recruited to a subset of the super-enhancer(SE)regions upstream of cell cycle-associated genes by OCT4,and increased their *** combining with promoter capture Hi-C data,we found that PCGF6 activates cell cycle genes by regulating SE-promoter interactions via 3D *** fin dings highlight a novel mechanism of PcG protein in regulating pluripotency,and provide a research basis for the therapeutic application of pluripotent stem cells.

关键词： PCGF6 Polycomb group super-enhancer 3D chromatin pluripotency

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Sperm chromatin condensation as an in vivo fertility biomarker in bulls:a flow cytometry approach

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Journal of Animal Science and Biotechnology 2022年第2期13卷 418-429页

作者： Marc Llavanera Jordi Ribas-Maynou Ariadna Delgado-Bermúdez Sandra Recuero Rodrigo Muiño Carlos OHidalgo Carolina Tamargo Sergi Bonet Yentel Mateo-Otero Marc Yeste Biotechnology of Animal and Human Reproduction(TechnoSperm) Faculty of SciencesUniversity of GironaC/Maria Aurèlia Campany69ES-17003 GironaSpain Department of Biology Unit of Cell BiologyFaculty of SciencesUniversity of GironaES-17003 GironaSpain Department of Animal Pathology Faculty of Veterinary MedicineUniversity of Santiago de CompostelaES-15705 LugoSpain Department of Animal Selection and Reproduction The Regional Agri-Food Research and Development Service of Asturias(SERIDA)E-33394 GijónSpain

Background:Genetic selection in cattle has been directed to increase milk ***,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the ***,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive *** humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male *** objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in *** study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein *** outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow *** quality parameters(morphology,viability,total and progressive motility)were also *** correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo *** morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile ***,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in ***,the percentage of viable sperm with poor chromatin condensation showed better accura

关键词： Bull chromatin Chromomycin A3 Condensation Fertility Flow cytometry Sperm

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Versatile functions of RNA m6A machinery on chromatin

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Journal of Molecular Cell Biology 2022年第3期14卷 3-11页

作者： Tanjing Song Suli Lv Neng Li Xuefeng Zhao Xianyun Ma Yingying Yan Weixia Wang Lidong Sun Department of Biochemistry and Molecular Biology School of Basic Medicineongji Medical College Huazhong University of Science and TechnologyWuhan 430030China

m6A,a conserved and abundant modification on RNA,regulates RNA processing and *** m6A machinery,including writers,erasers,and readers of m6A,is indispensable for m6A installation and ***,recent studies have revealed that m6A machinery can be recruited to chromatin by pleiotropic factors,including nascent RNA,transcription factors,regulatory RNA,histone modifications,and epigenetic ***,recruitment of m6A machinery can directly regulate chromatin biology,such as transcription,DNA damage repair,and DNA recombination beyond installation of m6A on nascent ***,we discuss recent evidence showing that m6A machinery is targeted to chromatin and the direct biological consequences along with the underlying mechanisms.

关键词： m6A chromatin RNA methylation histone modifications epigenetics

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