1.Host factor Moloney leukemia virus 10 (MOV10) protein inhibits replication of the xenotropic murine leukemia virus-related virus (XMRV).
Yue ZHANG ; Si-Qi HU ; Xiao-Jing PANG ; Jian LI ; Fei GUO
Chinese Journal of Virology 2014;30(5):514-520
We investigated inhibition of Moloney leukemia virus 10 (MOV10) upon xenotropic murine leukemia virus-related virus (XMRV) and made a preliminary study of the mechanism of action. Using transfection, infection, western blotting and real-time polymerase chain reaction, we found that MOV10 inhibited XMRV replication. Using MOV10 overexpressed in viral producer cells, MOV10 was shown to reduce the infectivity of XMRV. MOV10 could be incorporated into XMRV, suggesting that MOV10 could undergo encapsidation by XMRV during viral assembly. MOV10 could also restrict the DNA production of XMRV in target cells. We found that the putative RNA-helicase domain of MOV10 maintained most of its XMRV inhibition. These results suggest that MOV10 could be required during the retroviral lifecycle. Perturbation of MOV10 disrupts the generation of infectious viral particles, suggesting that MOV10 has broad antiretroviral activity. Hence, MOV10 could be actively involved in host defense against retroviral infection.
Humans
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Moloney murine leukemia virus
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physiology
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RNA Helicases
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physiology
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Virus Replication
2.Prokaryotic expression and purification of moloney murine leukemia virus reverse transcriptase and verification of the activity.
Xiansong WANG ; Xuemei MA ; Yi SUN
Chinese Journal of Biotechnology 2008;24(5):903-906
To produce the reverse transcriptase of moloney murine leukemia virus (MMLV-RT) through gene recombination, MMLV-rt gene was amplified by polymerase chain reaction (PCR) with specifically designed primers bearing restriction enzyme sites. Five mutation sites increasing the solution of the target protein were introduced through Site-directed mutation. After verification by sequencing, the gene was cloned into the expression vector pET15b to construct the recombinant plasmid pET15b-MMLV-rt. Purified MMLV-RT was obtained by affinity chromatography (Ni3+-NTA beads). Molecular weight and purity of MMLV-RT were analyzed with SDS-PAGE. Enzyme activity was characterized with RT-PCR. We successfully constructed the recombinant plasmid pET15b-MMLV-rt and obtained the MMLV-RT fusion protein with 6His on the N-terminus. Recombinant protein was purified through Ni3+-NTA beads based affinity chromatography, the purity of which was 96%. The Activity of the enzyme was high. MMLV-RT of 96% purity was obtained with the prokaryotic expression technique, which serves as the basis for mass production of this enzyme.
Animals
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Mice
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Moloney murine leukemia virus
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enzymology
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genetics
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RNA-Directed DNA Polymerase
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biosynthesis
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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metabolism
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Recombination, Genetic
3.B lymphoma Moloney murine leukemia virus insertion region 1: An oncogenic mediator in prostate cancer.
Qipeng LIU ; Qiaqia LI ; Sen ZHU ; Yang YI ; Qi CAO
Asian Journal of Andrology 2019;21(3):224-232
B lymphoma Moloney murine leukemia virus insertion region 1 (BMI1), a core member of polycomb repressive complex 1 (PRC1), has been intensely investigated in the field of cancer epigenetics for decades. Widely known as a critical regulator in cellular physiology, BMI1 is essential in self-renewal and differentiation in different lineages of stem cells. BMI1 also plays a significant role in cancer etiology for its involvement in pathological progress such as epithelial-mesenchymal transition (EMT) and cancer stem cell maintenance, propagation, and differentiation. Importantly, overexpression of BMI1 is predictive for drug resistance, tumor recurrence, and eventual therapy failure of various cancer subtypes, which renders the pharmacological targeting at BMI1 as a novel and promising therapeutic approach. The study on prostate cancer, a prevalent hormone-related cancer among men, has promoted enormous research advancements in cancer genetics and epigenetics. This review summarizes the role of BMI1 as an oncogenic and epigenetic regulator in tumor initiation, progression, and relapse of prostate cancer.
Animals
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Gene Expression Regulation, Neoplastic
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Humans
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Lymphoma, B-Cell/genetics*
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Male
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Mice
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Moloney murine leukemia virus/genetics*
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Mutagenesis, Insertional/genetics*
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Polycomb Repressive Complex 1/genetics*
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Prostatic Neoplasms/genetics*
4.Effect of Retinoic Acid on Growth and Transduced Tumor Necrosis Factor-alpha Gene Expression of Human Bladder Tumor Cell Lines.
Hyeon JEONG ; Sang Jin YOON ; Moon Ki JO ; Hae Won LEE ; Soo Woong KIM ; Eun Sik LEE ; Chong Wook LEE
Korean Journal of Urology 1997;38(3):229-234
INTRODUCTION AND OBJECTIVES: Retinoic acid (RA) is known as a potent chemopreventive agent in bladder tumor. Recently, RA has gained attention for up-regulation of transduced gene expression via long terminal repeat (LTR) transcriptional promotion. In this study, we investigated the possible dual effect of RA, growth inhibition and up-regulation of transduced gene expression which contains LTR promoter in human bladder carcinoma cell lines. MATERIALS AND METHODS: Human bladder carcinoma cell lines CY-24, J-82, HT-1197, ATCC) were transduced with Moloney murine leukemia virus containing cDNA of TNF-alpha. The growth of transduced and parent cell line was measured by tetrazolium based colorimetric assay (MTF). Transduced TNF-alpha gene expression was determined by ELISA method. RESULTS: TNF-alpha production was increased approximately twofold after treatment with RA (10 uM) in all three cell lines. This increase was dependent on RA concentration. RA treatment of transduced and parent cell line resulted in dose dependent inhibition of cell proliferation(up to 80% inhibitionwith 10 uM RA) in all parental and transduced cell lines. CONCLUSIONS: These results indicate that RA shows dual effect in cytokine gene transduced bladder carcinoma cells with retroviral vector containing LTR promoter and could be a supplement to the gene therapy of bladder cancer.
Cell Line*
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DNA, Complementary
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Enzyme-Linked Immunosorbent Assay
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Gene Expression*
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Genetic Therapy
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Humans*
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Moloney murine leukemia virus
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Parents
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Terminal Repeat Sequences
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Tretinoin*
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Tumor Necrosis Factor-alpha*
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Up-Regulation
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Urinary Bladder Neoplasms*
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Urinary Bladder*
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Zidovudine