MOLECULAR CELL
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2024
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IF 14.5
Huihui An,Yifan Hong,Yeek Teck Goh,Casslynn W.Q. Koh,Shahzina Kanwal,Yi Zhang,Zhaoqi Lu,Phoebe M.L. Yap,Suat Peng Neo,Chun-Ming Wong,Alice S.T. Wong,Yang Yu,Jessica Sook Yuin Ho,Jayantha Gunaratne,Wee Siong Sho Goh
N 6 ,2′-O-dimethyladenosine (m 6 Am) is an abundant mRNA modification that impacts multiple diseases, but its function remains controversial because the m 6 Am reader is unknown. Using quantitative proteomics, we identified transcriptional terminator premature cleavage factor II (PCF11) as a m 6 Am-specific reader in human cells. Direct quantification of mature versus nascent RNAs reveals that m 6 Am does not regulate mRNA stability but promotes nascent transcription. Mechanistically, m 6 Am functions by sequestering PCF11 away from proximal RNA polymerase II (RNA Pol II). This suppresses PCF11 from dissociating RNA Pol II near transcription start sites, thereby promoting full-length transcription of m 6 Am-modified RNAs. m 6 Am’s unique relationship with PCF11 means m 6 Am function is enhanced when PCF11 is reduced, which occurs during all- trans -retinoic-acid (ATRA)-induced neuroblastoma-differentiation therapy. Here, m 6 Am promotes expression of ATF3, which represses neuroblastoma biomarker MYCN. Depleting m 6 Am suppresses MYCN repression in ATRA-treated neuroblastoma and maintains their tumor-stem-like properties. Collectively, we characterize m 6 Am as an anti-terminator RNA modification that suppresses premature termination and modulates neuroblastoma’s therapeutic response.
研究领域:
分子生物学
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癌症生物学
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转录组学
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DOI
PubMed
高影响因子
NUCLEIC ACIDS RESEARCH
·
2022
·Vol. 50
·期 16
·9175-9189
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IF 19.16
Jia Ruirui,Lin Jiamei,You Jin,Li Shi,Shan Ge,Huang Chuan
Stress-responsive genes are lowly transcribed under normal conditions and robustly induced in response to stress. The significant difference between basal and induced transcription indicates that the general transcriptional machinery requires a mechanism to distinguish each transcription state. However, what factors specifically function in basal transcription remains poorly understood. Using a classic model stress-responsive gene (Drosophila MtnA), we found that knockdown of the DEAD-box helicase Hlc resulted in a significant transcription attenuation of MtnA under normal, but not stressed, conditions. Mechanistically, Hlc directly binds to the MtnA locus to maintain the accessibility of chromatin near the transcriptional start site, which allows the recruitment of RNA polymerase II and subsequent MtnA transcription. Using RNA-seq, we then identified plenty of additional stress-responsive genes whose basal transcription was reduced upon knockdown of Hlc. Taken together, these data suggest that Hlc-mediated basal transcription regulation is an essential and widespread mechanism for precise control of stress-responsive genes.
研究领域:
分子生物学
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转录调控
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基因表达
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DOI
PubMed
高影响因子
Colloid and Interface Science Communications
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2023
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IF 4.5
Peng Wang,Jiwen Geng,Chunyu Tan,Qiang Wei
Adhesive interface stiffness is capable of modulating cellular behavior and gene expression, yet the underlying mechanobiological mechanisms remain unclear. In this study, we investigated the effects of interface stiffness on gene expression pathways by hydrogels with divergent stiffness. Our results reveal that adhesive interface stiffness affects cytoplasmic mechanotranduction as well as nuclear mechanics, and ultimately regulating the subcellular localization of HDAC3. Further investigation unfolds that HDAC3 directly affects global acetylation levels within nucleus. And HDAC3-induced acetylation changes are regulated by myosin contraction, thereby portraying downstream gene expression. Additionally, our study indicates that the interface stiffness-mediated regulation of HDAC3 nuclear-cytoplasmic redistribution is dependent on CRM1, and inhibition of CRM1 impedes the nuclear export of HDAC3. In summary, our work provides an overview of how the subcellular localization of HDAC3 can be manipulated through the regulation of cell adhesion interface stiffness, thereby altering upstream RNA polymerase II activity and gene expression.
研究领域:
细胞生物学
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力学生物学
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分子生物学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2022
·Vol. 23
·期 7
·3833
·
IF 6.208
Mengling Huang,Abrar Ahmed,Wei Wang,Xue Wang,Cui Ma,Haowei Jiang,Wei Li,Lili Jing
Gene expression is tightly regulated during hematopoiesis. Recent studies have suggested that RNA polymerase II (Pol II) promoter proximal pausing, a temporary stalling downstream of the promoter region after initiation, plays a critical role in regulating the expression of various genes in metazoans. However, the function of proximal pausing in hematopoietic gene regulation remains largely unknown. The negative elongation factor (NELF) complex is a key factor important for this proximal pausing. Previous studies have suggested that NELF regulates granulocytic differentiation in vitro, but its in vivo function during hematopoiesis remains uncharacterized. Here, we generated the zebrafish mutant for one NELF complex subunit Nelfb using the CRISPR-Cas9 technology. We found that the loss ofnelfbselectively induced excessive granulocytic development during primitive and definitive hematopoiesis. The loss ofnelfbreduced hematopoietic progenitor cell formation and did not affect erythroid development. Moreover, the accelerated granulocytic differentiation and reduced progenitor cell development could be reversed by inhibiting Pol II elongation. Further experiments demonstrated that the other NELF complex subunits (Nelfa and Nelfe) played similar roles in controlling granulocytic development. Together, our studies suggested that NELF is critical in controlling the proper granulocytic development in vivo, and that promoter proximal pausing might help maintain the undifferentiated state of hematopoietic progenitor cells.Keywords:zebrafish;negative elongation factor;RNA polymerase II promoter proximal pausing;granulocytic development
研究领域:
分子生物学
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血液学
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发育生物学
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遗传学
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DOI
PubMed
高影响因子
BIOMEDICINE & PHARMACOTHERAPY
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2024
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IF 6.9
Junyu Wang,Chaojie Yang,Zhixin Liang,Junping Sun,Mingyue Zhang,Shaofu Qiu,Xinying Du,Xi He,Xiaoying Pang,Xidong Ma,Mei Xie,Xinjie Han,Ru Fan,Enlu Zhou,Hairong Yu,Danyang She,Hongbin Song,Jianxin Wang
Human adenovirus (HAdV) infection is a major cause of respiratory disease, yet no antiviral drugs have been approved for its treatment. Herein, we evaluated the antiviral and anti-inflammatory effects of cyclin-dependent protein kinase (CDK) inhibitor indirubin-3′-monoxime (IM) against HAdV infection in cells and a transgenic mouse model. After evaluating its cytotoxicity, cytopathic effect reduction, antiviral replication kinetics, and viral yield reduction assays were performed to assess the anti-HAdV activity of IM. Quantitative real-time polymerase chain reaction (qPCR), quantitative reverse transcription PCR (qRT-PCR), and western blotting were used to assess the effects of IM on HAdV DNA replication, transcription, and protein expression, respectively. IM significantly inhibited HAdV DNA replication as well as E1A and Hexon transcription, in addition to significantly suppressing the phosphorylation of the RNA polymerase II C-terminal domain (CTD). IM mitigated body weight loss, reduced viral burden, and lung injury, decreasing cytokine and chemokine secretion to a greater extent than cidofovir. Altogether, IM inhibits HAdV replication by downregulating CTD phosphorylation to suppress viral infection and corresponding innate immune reactions as a promising therapeutic agent.
研究领域:
病毒学
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药理学
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免疫学
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DOI
PubMed
高影响因子
ACS Nano
·
2021
·
IF 15.881
Chen-Song Huang,Qiong-Cong Xu,Chunlei Dai,Liying Wang,Yi-Chih Tien,Fuxi Li,Qiao Su,Xi-Tai Huang,Jun Wu,Wei Zhao,Xiao-Yu Yin
Gallbladder cancer (GBC) is the most aggressive malignancy of the biliary tract cancer, and there is a lack of effective treatment. Here, we developed a nanoparticle platform (8P4 NP) that can deliver THZ1, a cyclin-dependent kinase 7 (CDK7) inhibitor, to treat GBC. Analysis of datasets demonstrated that CDK7 was positively correlated with poor prognosis. CDK7 inhibition suppressed cell proliferation, induced apoptosis, and caused cell cycle block in GBC cells. THZ1 downregulated CDK7-mediated phosphorylation of RNA polymerase II (RNAPII), resulting in a significant downregulation of transcriptional programs, with a preferential repression of oncogenic transcription factors. To improve the tumor targeting efficiency of THZ1, 8P4 NPs were prepared and assembled with THZ1 to form THZ1@8P4 NPs. Compared with free THZ1, THZ1@8P4 NPs showed more advantages in prolonging blood circulation, escaping from lysosomes and increasing cellular uptake. Importantly, THZ1@8P4 NPs demonstrated a more significant inhibition effect on GBC cells than free THZ1 in vitro. In addition, THZ1@8P4 NPs could efficiently deliver THZ1 to tumor sites in a patient-derived xenograft model of early recurrence, leading to tumor regression and transcriptional inhibition with minimal toxicity. In summary, we conclude that THZ1@8P4 NPs provide a potent therapeutic strategy that targets CDK7-mediated transcriptional addiction in GBC.
研究领域:
肿瘤学
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药理学
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纳米技术
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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