المؤلفون: Chen, Xinyan, Raiff, Anat, Li, Shanshan, Guo, Qiong, Zhang, Jiahai, Zhou, Hualin, Timms, Richard T, Yao, Xuebiao, Elledge, Stephen J, Koren, Itay, Zhang, Kaiming, Xu, Chao

مصطلحات موضوعية: Humans, Ubiquitination, Cryoelectron Microscopy, Ubiquitin-Protein Ligases, NEDD8 Protein, Proline, Protein Multimerization, HEK293 Cells, Protein Binding, Substrate Specificity, Cell Cycle Proteins, Models, Molecular, Cullin Proteins, Degrons, Receptors, Interleukin-17

الوصف: Acknowledgements: We thank Dr. Yong-Xiang Gao and the Cryo-EM Center at the University of Science and Technology of China for technical support with cryo-EM data collection; Prof. Yarden Opatowsky and Julia Guez-Haddad for gel filtration. This work is supported by the National Key R&D Program of China 2022YFA1303100, the National Natural Science Foundation of China (22137007 and 92253301), the Research Funds of Center for Advanced Interdisciplinary Science and Biomedicine of IHM (QYPY20230022), the Ministry of Science and Technology of China (2022YFC2303700 and 2022YFA1302700), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB0490000), the Center for Advanced Interdisciplinary Science and Biomedicine of IHM (QYPY20220019), and the Fundamental Research Funds for the Central Universities (WK9100000032, WK9100000044, and WK9100000027). C.X. is also supported by the Major/Innovative Program of Development Foundation of Hefei Center for Physical Science and Technology (2021HSC-CIP014); I.K. is supported by the European Research Council (ERC-2020-STG 947709), Israel Science Foundation (ISF Grants No. 2380/21 and 3096/21), Alon Fellowship and Applebaum Foundation. R.T.T. is a Sir Henry Wellcome Postdoctoral Fellow (201387/Z/16/Z) and a Pemberton-Trinity Fellow. ; Funder: the National Key R&D Program of China 2022YFA1303100 ; Funder: the Ministry of Science and Technology of China (2022YFC2303700 and 2022YFA1302700) ; Funder: the Ministry of Science and Technology of China (2022YFC2303700 and 2022YFA1302700),the Strategic Priority Research Program of the Chinese Academy of Sciences XDB0490000, the Center for Advanced Interdisciplinary Science Biomedicine of IHM QYPY20220019 ; The E3 ligase-degron interaction determines the specificity of the ubiquitin‒proteasome system. We recently discovered that FEM1B, a substrate receptor of Cullin 2-RING ligase (CRL2), recognizes C-degrons containing a C-terminal proline. By solving several cryo-EM structures of CRL2FEM1B bound to different ...

وصف الملف: text/xml; application/pdf; application/zip

العلاقة: https://www.repository.cam.ac.uk/handle/1810/367693Test

الإتاحة: https://www.repository.cam.ac.uk/handle/1810/367693Test

المؤلفون: Mateja Jagić, Tamara Vuk, Andreja Škiljaica, Lucija Markulin, Vedrana Vičić Bočkor, Mirta Tokić, Karlo Miškec, Genadij Razdorov, Siniša Habazin, Marko Šoštar, Igor Weber, Nataša Bauer, Dunja Leljak Levanić

المصدر: Plant Cell Reports. 41:2139-2157

مصطلحات موضوعية: Homeodomain Proteins, MATH-BTB, DNA methylation, RdDM, DDR complex, Protein interactions, Arabidopsis thaliana, Arabidopsis Proteins, Arabidopsis, RNA, Plant Science, General Medicine, DNA Methylation, Cullin Proteins, Agronomy and Crop Science, Transcription Factors

الوصف: BPM1 interacts with components of the DDR complex and stimulates DNA methylation at CHH sites, suggesting its involvement in the RdDM methylation pathway. The best-known function of MATH-BTB proteins, including Arabidopsis BPM proteins, is their role as substrate-specific adaptors of CUL3-based E3 ligases in the ubiquitin-proteasome pathway. This paper reports a new CUL3-independent role of BPM1 in RNA-directed DNA methylation (RdDM). Using quantitative and qualitative Y2H, pull down, microscale thermophoresis and FRET-FLIM, we demonstrate that BPM1 interacts with DMS3 and RDM1, components of the chromatin remodeling DDR complex involved in the recruitment of the RdDM methylation machinery. All three proteins colocalized predominantly in the nucleus. The MATH domain, which specifically binds proteins destined for degradation, was not essential for interactions with DMS3 and RDM1. In plants overexpressing BPM1, endogenous DMS3 protein levels were stable, indicating that BPM1 does not induce proteasomal degradation. In RDM1-overexpressing plants, RDM1 was not ubiquitinated. Together, these results suggest that BPM1 does not mediate the degradation of DMS3 and RDM1. Additionally, overexpression of BPM1 caused increased global methylation levels as well as CHH methylation in promoters of two RdDM-regulated genes, FWA and CML41. Overall, BPM1 seems to have a stimulating effect on RdDM activity, and this role appears to be unrelated to its known function as a Cul3-based E3 ligase adaptor.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ec3a9674c274f588ccf09ff4c76e286bTest
https://doi.org/10.1007/s00299-022-02911-9Test

المؤلفون: Yan Lin, Qian Li, Xiaofeng Jin

المصدر: Molecular Biology Reports. 49:9813-9824

مصطلحات موضوعية: Ubiquitin, Pseudohypoaldosteronism, Microfilament Proteins, Genetics, Humans, General Medicine, Protein Serine-Threonine Kinases, Carrier Proteins, Cullin Proteins, Molecular Biology, Adaptor Proteins, Signal Transducing

الوصف: Kelch-like protein 3 (KLHL3) is a substrate adaptor of Cullin3-RING ubiquitin ligase (CRL3), and KLHL3-CUL3 complex plays a vital role in the ubiquitination of specific substrates. Mutations and abnormal post-translational modifications of KLHL3-CUL3 affect substrate ubiquitination and may related to the pathogenesis of Gordon syndrome (GS), Primary Hyperparathyroidism (PHPT), Diabetes Mellitus (DM), Congenital Heart Disease (CHD), Pre-eclampsia (PE) and even cancers. Therefore, it is essential to understand the function and molecular mechanisms of KLHL3-CUL3 for the treatment of related diseases. In this review, we summary the structure and function of KLHL3-CUL3, the effect of KLHL3-CUL3 mutations and aberrant modifications in GS, PHPT, DM, CHD and PE. Moreover, we noted a possible role of KLHL3-CUL3 in carcinogenesis and provided ideas for targeting KLHL3-CUL3 for related disease treatment.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::91471cbf6b68e8df3678e8404779cec3Test
https://doi.org/10.1007/s11033-022-07487-xTest

المؤلفون: Tiantian Xu, Qisheng Ma, Yanan Li, Qing Yu, Peichen Pan, Yawen Zheng, Zhijian Li, Xiufang Xiong, Tingjun Hou, Bin Yu, Hongmin Liu, Yi Sun

المصدر: Signal Transduction and Targeted Therapy. 7

مصطلحات موضوعية: G2 Phase Cell Cycle Checkpoints, DNA Adducts, Cancer Research, Lung Neoplasms, Pyrimidines, Cell Line, Tumor, Ubiquitin-Protein Ligases, Ubiquitin-Conjugating Enzymes, Genetics, Humans, Apoptosis, Cyclopentanes, Cullin Proteins

الوصف: Protein neddylation is catalyzed by a neddylation activating enzyme (NAE, E1), an E2 conjugating enzyme, and an E3 ligase. In various types of human cancers, the neddylation pathway is abnormally activated. Our previous study validated that the neddylation E2 UBE2F is a promising therapeutic target in lung cancer. Although the NAE inhibitor MLN4924/pevonedistat is currently under clinical investigation as an anti-cancer agent, there are no small molecules available that selectively target UBE2F. Here, we report, for the first time, the discovery, via structure-based virtual screen and chemical optimization, of such a small molecule, designated as HA-9104. HA-9104 binds to UBE2F, reduces its protein levels, and consequently inhibits cullin-5 neddylation. Blockage of cullin-5 neddylation inactivates cullin-RING ligase-5 (CRL5) activity, leading to accumulation of the CRL5 substrate, NOXA, to induce apoptosis. Moreover, HA-9104 appears to form the DNA adduct via its 7-azaindole group to induce DNA damage and G2/M arrest. Biologically, HA-9104 effectively suppresses the growth and survival of lung cancer cells and confers radiosensitization in both in vitro cell culture and in vivo xenograft tumor models. In summary, we discovered a small molecule, designated HA-9104, that targets the UBE2F-CRL5 axis with anti-cancer activity alone or in combination with radiation.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f650be5d325527678443c849caecc3a4Test
https://doi.org/10.1038/s41392-022-01182-wTest

المؤلفون: Jian, Shen, Youqi, Li, Menghao, Li, Zhiming, Li, Huantang, Deng, Xiongwei, Xie, Jinguang, Liu

المصدر: BioMedical Engineering OnLine. 21

مصطلحات موضوعية: Radiological and Ultrasound Technology, Angiotensin II, Biomedical Engineering, Gene Expression, General Medicine, Cullin Proteins, Muscle, Smooth, Vascular, Biomaterials, Mice, Hypertension, Animals, Hedgehog Proteins, Radiology, Nuclear Medicine and imaging, Reactive Oxygen Species, Cells, Cultured, Cell Proliferation, Signal Transduction

الوصف: Background Hypertension is known as a major factor for global mortality. We aimed to investigate the role of Cullin3 (CUL3) in the regulation of hypertension. Material and methods Human vascular smooth muscle cells (VSMCs) were treated with Angiotensin II (Ang II) to establish a hypertension in vitro model. Cell viability was detected by a cell counting kit-8 (CCK-8) assay. The content of reactive oxygen species (ROS) was evaluated by kit. Transwell assay and TUNEL staining were, respectively, used to assess cell migration and apoptosis. Additionally, the expression of sonic hedgehog (SHH) signaling-related proteins (SHH, smoothened homolog (Smo) and glioblastoma (Gli)) and CUL3 was tested with western blotting. Following treatment with Cyclopamine (Cycl), an inhibitor of SHH signaling, in Ang II-induced VSMCs, cell viability, migration, apoptosis and ROS content were determined again. Then, VSMCs were transfected with CUL3 plasmid or/and treated with sonic hedgehog signaling agonist (SAG) to explore the impacts on Ang II-induced VSMCs damage. In vivo, a hypertensive mouse model was established. Systolic blood pressure and diastolic blood pressure were determined. The histopathologic changes of abdominal aortic tissues were examined using H&E staining. The expression of SHH, Smo, Gli and CUL3 was tested with western blotting. Results Significantly increased proliferation, migration and apoptosis of VSMCs were observed after Ang II exposure. Moreover, Ang II induced upregulated SHH, Smo and Gli expression, whereas limited increase in CUL3 expression was observed. The content of ROS in Ang II-stimulated VSMCs presented the same results. Following Cycl treatment, the high levels of proliferation and migration in Ang II-treated VSMCs were notably remedied while the apoptosis and ROS concentration were further increased. Moreover, Cycl downregulated SHH, Smo, Gli and CUL3 expression. Above-mentioned changes caused by Ang II were reversed following SAG addition. Indeed, SAG treatment combined with restoration of CUL3 expression inhibited proliferation, migration, apoptosis and ROS level in Ang II-stimulated VSMCs. In vivo, SAG aggravated the histopathological changes of the aorta and with a worse tendency after both SAG intervention and CUL3 silencing. By contrast, SAG treatment and rebound in CUL3 expression alleviated the vascular damage. Conclusions Collectively, restoration of CUL3 gene expression protected against hypertension through enhancing the effects of SHH activation in inhibition of apoptosis and oxidative stress for hypertension and alleviating the dysfunction of VSMCs.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::76eecefaebd878e5a501dd553e2fe10fTest
https://doi.org/10.1186/s12938-022-01002-wTest

المؤلفون: Di Wu, Haomin Li, Mingwei Liu, Jun Qin, Yi Sun

المصدر: Nature Communications. 13

مصطلحات موضوعية: Mice, Multidisciplinary, Ubiquitin-Conjugating Enzymes, Ubiquitination, Animals, General Physics and Astronomy, General Chemistry, Carrier Proteins, Cullin Proteins, T-Lymphocytes, Regulatory, General Biochemistry, Genetics and Molecular Biology

الوصف: Neddylation-mediated activation of Cullin-RING E3 Ligases (CRLs) are necessary for the degradation of specific immune regulatory proteins. However, little is known about how these processes govern the function of regulatory T (Treg) cells. Here we show that mice with Treg cell-specific deletion of Rbx1, a dual E3 for both neddylation and ubiquitylation by CRLs, develop an early-onset fatal inflammatory disorder, characterized by disrupted Treg cell homeostasis and suppressive functions. Specifically, Rbx1 is essential for the maintenance of an effector Treg cell subpopulation, and regulates several inflammatory pathways. Similar but less severe phenotypes are observed in mice having Ube2m, a neddylation E2 conjugation enzyme, deleted in their Treg cells. Interestingly, Treg-specific deletion of Rbx2/Sag or Ube2f, components of a similar but distinct neddylation-CRL complex, yields no obvious phenotype. Thus, our work demonstrates that the Ube2m-Rbx1 axis is specifically required for intrinsic regulatory processes in Treg cells; and that Rbx1 might also play Ube2m-independent roles in maintaining the fitness of Treg cells, suggesting a layer of complexity in neddylation-dependent activation of CRLs.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1aa2153d631d8ba0f74023dedf301bc0Test
https://doi.org/10.1038/s41467-022-30707-8Test

المؤلفون: Xiuli Zhang, Tong Ji, Ruihong Zhang, Ping Liu, Donghe Li, Xingyu Lv, Ruibao Ren, Mingzhu Liu, Zhizhou Xia

المصدر: Frontiers of Medicine. 15:302-312

مصطلحات موضوعية: Male, 0301 basic medicine, Ubiquitin-Protein Ligases, Motility, Biology, Mice, 03 medical and health sciences, 0302 clinical medicine, Animals, Reproductive system, Spermatogenesis, Acrosome, Receptor, Mice, Knockout, Factor VIII, General Medicine, Cullin Proteins, Sperm, Cell biology, Ubiquitin ligase, DNA-Binding Proteins, 030104 developmental biology, Knockout mouse, biology.protein, 030217 neurology & neurosurgery

الوصف: Cullin-RING E3 ubiquitin ligase (CRL)-4 is a member of the large CRL family in eukaryotes. It plays important roles in a wide range of cellular processes, organismal development, and physiological and pathological conditions. DDB1- and CUL4-associated factor 8 (DCAF8) is a WD40 repeat-containing protein, which serves as a substrate receptor for CRL4. The physiological role of DCAF8 is unknown. In this study, we constructed Dcaf8 knockout mice. Homozygous mice were viable with no noticeable abnormalities. However, the fertility of Dcaf8-deficient male mice was markedly impaired, consistent with the high expression of DCAF8 in adult mouse testis. Sperm movement characteristics, including progressive motility, path velocity, progressive velocity, and track speed, were significantly lower in Dcaf8 knockout mice than in wild-type (WT) mice. However, the total motility was similar between WT and Dcaf8 knockout sperm. More than 40% of spermatids in Dcaf8 knockout mice showed pronounced morphological abnormalities with typical bent head malformation. The acrosome and nucleus of Dcaf8 knockout sperm looked similar to those of WT sperm. In vitro tests showed that the fertilization rate of Dcaf8 knockout mice was significantly reduced. The results demonstrated that DCAF8 plays a critical role in spermatogenesis, and DCAF8 is a key component of CRL4 function in the reproductive system.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5bb52e52a533c70042327536b77136b1Test
https://doi.org/10.1007/s11684-021-0851-8Test

المؤلفون: Rahul Satija, William M. Mauck, Bernadette Bracken, Andrew Butler, Efthymia Papalexi, Eleni P. Mimitou, Yuhan Hao, Hans-Hermann Wessels, Samantha D. Foster, Bertrand Z. Yeung, Peter Smibert

المصدر: Nature genetics

مصطلحات موضوعية: NF-E2-Related Factor 2, THP-1 Cells, Cell, Cell Cycle Proteins, Computational biology, Signal-To-Noise Ratio, Biology, B7-H1 Antigen, Article, 03 medical and health sciences, 0302 clinical medicine, Immune system, Downregulation and upregulation, Interferon, Genetics, medicine, Humans, CRISPR, Clustered Regularly Interspaced Short Palindromic Repeats, Receptors, Interferon, 030304 developmental biology, 0303 health sciences, Messenger RNA, Kelch-Like ECH-Associated Protein 1, Mechanism (biology), Reproducibility of Results, Cullin Proteins, Immune Checkpoint Proteins, Programmed Cell Death 1 Ligand 2 Protein, KEAP1, medicine.anatomical_structure, B7-2 Antigen, Single-Cell Analysis, 030217 neurology & neurosurgery, Transcription Factors, medicine.drug

الوصف: The expression of inhibitory immune checkpoint molecules, such as programmed death-ligand (PD-L)1, is frequently observed in human cancers and can lead to the suppression of T cell-mediated immune responses. Here, we apply expanded CRISPR-compatible (EC)CITE-seq, a technology that combines pooled CRISPR screens with single-cell mRNA and surface protein measurements, to explore the molecular networks that regulate PD-L1 expression. We also develop a computational framework, mixscape, that substantially improves the signal-to-noise ratio in single-cell perturbation screens by identifying and removing confounding sources of variation. Applying these tools, we identify and validate regulators of PD-L1 and leverage our multimodal data to identify both transcriptional and post-transcriptional modes of regulation. Specifically, we discover that the Kelch-like protein KEAP1 and the transcriptional activator NRF2 mediate the upregulation of PD-L1 after interferon (IFN)-γ stimulation. Our results identify a new mechanism for the regulation of immune checkpoints and present a powerful analytical framework for the analysis of multimodal single-cell perturbation screens.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::302ec219a1bd39f2dc8c2191eb1ef9fbTest
https://doi.org/10.1038/s41588-021-00778-2Test

المؤلفون: Yunshan Wang, Guangwei Wei, Jian-Hua Mao, Maoxiao Feng, Pengju Zhang, Huaizhi Wang, Zhongxi Zhao, Lei Bi

المصدر: Oncogene
Oncogene, vol 40, iss 11

مصطلحات موضوعية: 0301 basic medicine, Genome instability, Cancer Research, DNA End-Joining Repair, Ubiquitylation, DNA Repair, Carcinogenesis, DNA-Activated Protein Kinase, medicine.disease_cause, Malignant transformation, 0302 clinical medicine, 2.1 Biological and endogenous factors, Aetiology, DNA-PKcs, Cancer, chemistry.chemical_classification, Tumor, biology, Nuclear Proteins, Cullin Proteins, Ubiquitin ligase, 030220 oncology & carcinogenesis, DNA damage, Clinical Sciences, Oncology and Carcinogenesis, Article, Genomic Instability, Cell Line, 03 medical and health sciences, Cell Line, Tumor, Genetics, medicine, Humans, Oncology & Carcinogenesis, Non-homologous-end joining, Molecular Biology, Cell Proliferation, Cell Nucleus, DNA ligase, fungi, enzymes and coenzymes (carbohydrates), 030104 developmental biology, chemistry, biology.protein, Cancer research, CUL4A, Precancerous Conditions, DNA Damage

الوصف: Genomic instability induced by DNA damage and improper DNA damage repair is one of the main causes of malignant transformation and tumorigenesis. DNA double strand breaks (DSBs) are the most detrimental form of DNA damage, and nonhomologous end-joining (NHEJ) mechanisms play dominant and priority roles in initiating DSB repair. A well-studied oncogene, the ubiquitin ligase Cullin 4A (CUL4A), is reported to be recruited to DSB sites in genomic DNA, but whether it regulates NHEJ mechanisms of DSB repair is unclear. Here, we discovered that the CUL4A-DTL ligase complex targeted the DNA-PKcs protein in the NHEJ repair pathway for nuclear degradation. Overexpression of either CUL4A or DTL reduced NHEJ repair efficiency and subsequently increased the accumulation of DSBs. Moreover, we demonstrated that overexpression of either CUL4A or DTL in normal cells led to genomic instability and malignant proliferation. Consistent with the in vitro findings, in human precancerous lesions, CUL4A expression gradually increased with increasing malignant tendency and was negatively correlated with DNA-PKcs and positively correlated with γ-H2AX expression. Collectively, this study provided strong evidence that the CUL4A-DTL axis increases genomic instability and enhances the subsequent malignant transformation of normal cells by inhibiting NHEJ repair. These results also suggested that CUL4A may be a prognostic marker of precancerous lesions and a potential therapeutic target in cancer.

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الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0e4a49572ba3581c0dc4a5ac7eea8728Test
https://doi.org/10.1038/s41388-021-01690-zTest

المؤلفون: Murthy, Meena, Kurz, Thimo, O'Shaughnessy, Kevin M

مصطلحات موضوعية: Hyperkalemia, Hypertension, NCC, Proteasome, SPAK/OSR1 phosphorylation, Ubiquitin-E3 ligase complex, WNK kinases, Animals, Blood Pressure, Cullin Proteins, Humans, Neovascularization, Physiologic, Protein Serine-Threonine Kinases, Pseudohypoaldosteronism, Receptors, Drug, Signal Transduction, Sodium Chloride Symporters, Sodium-Potassium-Chloride Symporters

الوصف: Hypertension (high blood pressure) is a major public health problem affecting more than a billion people worldwide with complications, including stroke, heart failure and kidney failure. The regulation of blood pressure is multifactorial reflecting genetic susceptibility, in utero environment and external factors such as obesity and salt intake. In keeping with Arthur Guyton's hypothesis, the kidney plays a key role in blood pressure control and data from clinical studies; physiology and genetics have shown that hypertension is driven a failure of the kidney to excrete excess salt at normal levels of blood pressure. There is a number of rare Mendelian blood pressure syndromes, which have shed light on the molecular mechanisms involved in dysregulated ion transport in the distal kidney. One in particular is Familial hyperkalemic hypertension (FHHt), an autosomal dominant monogenic form of hypertension characterised by high blood pressure, hyperkalemia, hyperchloremic metabolic acidosis, and hypercalciuria. The clinical signs of FHHt are treated by low doses of thiazide diuretic, and it mirrors Gitelman syndrome which features the inverse phenotype of hypotension, hypokalemic metabolic alkalosis, and hypocalciuria. Gitelman syndrome is caused by loss of function mutations in the thiazide-sensitive Na/Cl cotransporter (NCC); however, FHHt patients do not have mutations in the SCL12A3 locus encoding NCC. Instead, mutations have been identified in genes that have revealed a key signalling pathway that regulates NCC and several other key transporters and ion channels in the kidney that are critical for BP regulation. This is the WNK kinase signalling pathway that is the subject of this review. ; KMO and MM would like to thank the British Heart Foundation for support in some of their work cited in this review (PG/13/89/30577).

وصف الملف: Print-Electronic; application/pdf

العلاقة: https://www.repository.cam.ac.uk/handle/1810/262011Test

الإتاحة: https://doi.org/10.17863/CAM.7250Test
https://www.repository.cam.ac.uk/handle/1810/262011Test