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Minichromosome modification

Yan Zhang, Song He, Jin-Jun Guo, Hong Peng, Jia-Hao Fan, Qing-Ling Li
BACKGROUND AND AIM: The HBV covalently closed circular DNA (cccDNA) is organized into a minichromosome in the nuclei of infected hepatocytes through interactions with histone and nonhistone proteins. Retinoid X receptor α (RXRα), a liver-enriched nuclear receptor, participates in regulation of HBV replication and transcription through modulation of HBV enhancer 1 and core promoter activity. MATERIAL AND METHODS: This study investigated RXRα involvement in HBV cccDNA epigenetic modifications...
August 1, 2017: Annals of Hepatology
Mingfeng Cao, Meirong Gao, Carmen Lorena Lopez-Garcia, Yutong Wu, Arun Somwarpet Seetharam, Andrew Josef Severin, Zengyi Shao
Many nonconventional yeast species have highly desirable features that are not possessed by model yeasts, despite that significant technology hurdles to effectively manipulate them lay in front. Scheffersomyces stipitis is one of the most important exemplary nonconventional yeasts in biorenewables industry, which has a high native xylose utilization capacity. Recent study suggested its much better potential than Saccharomyces cerevisiae as a well-suited microbial biomanufacturing platform for producing high-value compounds derived from shikimate pathway, many of which are associated with potent nutraceutical or pharmaceutical properties...
August 18, 2017: ACS Synthetic Biology
Kathrin Deuschle, Gabi Kepp, Holger Jeske
Geminiviral minichromosomes were purified to explore epigenetic modifications. The levels of methylation in their covalently closed circular DNA were examined with the help of methylation-dependent restriction (MdR). DNA with 12 superhelical turns was preferentially modified, indicating minichromosomes with 12 nucleosomes leaving an open gap. MdR digestion yielded a specific product of genomic length, which was cloned and Sanger-sequenced, or amplified following ligation-mediated rolling circle amplification and deep-sequenced (circomics)...
December 2016: Virology
Esther Adriana Ceniceros-Ojeda, Edgar Antonio Rodríguez-Negrete, Rafael Francisco Rivera-Bustamante
UNLABELLED: Geminiviruses are important plant pathogens characterized by circular, single-stranded DNA (ssDNA) genomes. However, in the nuclei of infected cells, viral double-stranded DNA (dsDNA) associates with host histones to form a minichromosome. In phloem-limited geminiviruses, the characterization of viral minichromosomes is hindered by the low concentration of recovered complexes due to the small number of infected cells. Nevertheless, geminiviruses are both inducers and targets of the host posttranscriptional gene silencing (PTGS) and transcriptional gene silencing (TGS) machinery...
April 2016: Journal of Virology
Gianna Aurora Palumbo, Cecilia Scisciani, Natalia Pediconi, Leonardo Lupacchini, Dulce Alfalate, Francesca Guerrieri, Ludovica Calvo, Debora Salerno, Silvia Di Cocco, Massimo Levrero, Laura Belloni
The HBV covalently closed circular DNA (cccDNA) is organized as a mini-chromosome in the nuclei of infected hepatocytes by histone and non-histone proteins. Transcription from the cccDNA of the RNA replicative intermediate termed pre-genome (pgRNA), is the critical step for genome amplification and ultimately determines the rate of HBV replication. Multiple evidences suggest that cccDNA epigenetic modifications, such as histone modifications and DNA methylation, participate in regulating the transcriptional activity of the HBV cccDNA...
2015: PloS One
Chiung-Yao Fang, Cheng-Huang Shen, Meilin Wang, Pei-Lain Chen, Michael W Y Chan, Pang-Hung Hsu, Deching Chang
During polyomavirus infection, the viral DNA adopts histones from host cells and forms minichromosomes as an important part of the viral life cycle. However, the detailed mechanisms of this histone incorporation remain unclear. Here, we profiled the histone posttranslational modifications (PTMs) in BKPyV minichromosomes and in the chromatin of BKPyV host cells. Through Triton-acetic acid-urea (TAU)-PAGE separation followed by nanoflow liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis, we identified different kinds of PTMs on histones from BKPyV minichromosomes and from host cells...
September 2015: Virology
Emma L Hesketh, Richard P Parker-Manuel, Yuriy Chaban, Rabab Satti, Dawn Coverley, Elena V Orlova, James P J Chong
ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in higher eukaryotes such as Drosophila, MCM-associated DNA helicase activity has been observed only in the context of a co-purified Cdc45-MCM-GINS complex. Here, we describe the production of the recombinant human MCM (hMCM) complex in Escherichia coli. This protein displays ATP hydrolysis activity and is capable of unwinding duplex DNA...
March 20, 2015: Journal of Biological Chemistry
Björn Krenz, Kathrin Deuschle, Tobias Deigner, Sigrid Unseld, Gabi Kepp, Christina Wege, Tatjana Kleinow, Holger Jeske
UNLABELLED: The C2/AC2 genes of monopartite/bipartite geminiviruses of the genera Begomovirus and Curtovirus encode important pathogenicity factors with multiple functions described so far. A novel function of Abutilon mosaic virus (AbMV) AC2 as a replication brake is described, utilizing transgenic plants with dimeric inserts of DNA B or with a reporter construct to express green fluorescent protein (GFP). Their replicational release upon AbMV superinfection or the individual and combined expression of epitope-tagged AbMV AC1, AC2, and AC3 was studied...
April 2015: Journal of Virology
Daniel Latreille, Lisa Bluy, Monsef Benkirane, Rosemary E Kiernan
The epigenome is defined as a type of information that can be transmitted independently of the DNA sequence, at the chromatin level, through post-translational modifications present on histone tails. Recent advances in the identification of histone 3 variants suggest a new model of information transmission through deposition of specific histone variants. To date, several non-centromeric histone 3 variants have been identified in mammals. Despite protein sequence similarity, specific deposition complexes have been characterized for both histone 3...
April 2014: Nucleic Acids Research
Hasan Yardimci, Johannes C Walter
In G1, two copies of the MCM2-7 helicase are recruited to each origin of replication. Whereas recruitment of the first MCM2-7 is likely to be analogous to the loading of sliding clamps around DNA, how the second MCM2-7 complex is recruited is highly contentious. Here, we argue that MCM2-7 loading involves specific modifications to the clamp-loading reaction and propose that the first and second MCM2-7 molecules are loaded via similar mechanisms.
January 2014: Nature Structural & Molecular Biology
S Etemad-Moghadam, S Fouladdel, E Azizi, M Alaeddini
PURPOSE: Aberrant proliferation is an essential feature of cancer cells, which can be caused by alterations in components of the cell cycle, such as minichromosome maintenance protein-3 (MCM3) and Ki-67. Doxorubicin is a cytotoxic/cytostatic anticancer agent commonly used in chemotherapy. We investigated the effect of this drug on MCM3 and Ki-67 in the KB cell line, which is considered a subline of HeLa cell line. METHODS: KB cells were treated with doxorubicin and its effect on apoptosis, mRNA levels and protein expression of MCM3 and Ki-67 was determined by flow cytometry (annexin V-FITC/PI assay), quantitative real-time RT-PCR (qRT-PCR) and immunocytochemistry, respectively...
October 2013: Journal of B.U.ON.: Official Journal of the Balkan Union of Oncology
Xiaoyong Zhang, Jinlin Hou, Mengji Lu
The hepatitis B virus (HBV) genome forms a covalently closed circular DNA (cccDNA) minichromosome that persists in the nucleus of virus-infected hepatocytes. HBV cccDNA serves as the template for viral mRNA synthesis and is subject to epigenetic regulation by several mechanisms, including DNA methylation and histone acetylation. Recently, microRNAs (miRNAs), a class of small non-coding RNAs, were also directly connected to the epigenetic machinery through a regulatory loop. Epigenetic modifications have been shown to affect miRNA expression, and a sub-group of miRNAs (defined as epi-miRNAs) can directly target effectors of the epigenetic machinery...
October 14, 2013: Frontiers in Genetics
Fei Liu, Matthew Campagna, Yonghe Qi, Xuesen Zhao, Fang Guo, Chunxiao Xu, Sichen Li, Wenhui Li, Timothy M Block, Jinhong Chang, Ju-Tao Guo
Covalently closed circular DNA (cccDNA) of hepadnaviruses exists as an episomal minichromosome in the nucleus of infected hepatocyte and serves as the transcriptional template for viral mRNA synthesis. Elimination of cccDNA is the prerequisite for either a therapeutic cure or immunological resolution of HBV infection. Although accumulating evidence suggests that inflammatory cytokines-mediated cure of virally infected hepatocytes does occur and plays an essential role in the resolution of an acute HBV infection, the molecular mechanism by which the cytokines eliminate cccDNA and/or suppress its transcription remains elusive...
September 2013: PLoS Pathogens
Isaac B Hilton, Jeremy M Simon, Jason D Lieb, Ian J Davis, Blossom Damania, Dirk P Dittmer
Kaposi's sarcoma-associated herpesvirus (KSHV) is an oncogenic gammaherpesvirus which establishes latent infection in endothelial and B cells, as well as in primary effusion lymphoma (PEL). During latency, the viral genome exists as a circular DNA minichromosome (episome) and is packaged into chromatin analogous to human chromosomes. Only a small subset of promoters, those which drive latent RNAs, are active in latent episomes. In general, nucleosome depletion ("open chromatin") is a hallmark of eukaryotic regulatory elements such as promoters and transcriptional enhancers or insulators...
November 2013: Journal of Virology
Les Kallestad, Emily Woods, Kendra Christensen, Amanda Gefroh, Lata Balakrishnan, Barry Milavetz
Simian virus 40 (SV40) early transcription is repressed when the product of early transcription, T-antigen, binds to its cognate regulatory sequence, Site I, in the promoter of the SV40 minichromosome. Because SV40 minichromosomes undergo replication and transcription potentially repression could occur during active transcription or during DNA replication. Since repression is frequently epigenetically marked by the introduction of specific forms of methylated histone H3, we characterized the methylation of H3 tails during transcription and replication in wild-type SV40 minichromosomes and mutant minichromosomes which did not repress T-antigen expression...
2013: Frontiers in Genetics
Kevin Y Kim, Steve B Huerta, Chie Izumiya, Don-Hong Wang, Anthony Martinez, Bogdan Shevchenko, Hsing-Jien Kung, Mel Campbell, Yoshihiro Izumiya
Kaposi's sarcoma-associated herpesvirus (KSHV) latent genomes are tethered to host histones to form a minichromosome also known as an "episome." Histones, which are core components of chromatin, are heavily modified by various histone-targeting enzymes. Posttranslational modifications of histones significantly influence accessibility of transcriptional factors and thus have profound effects on gene expression. Recent studies showed that epigenetic marks on the KSHV episome are well organized, exemplified by the absence of histone H3 lysine 9 (H3K9) methylation, a heterochromatic histone mark, from immediate early and latent gene promoters in naturally infected cells...
June 2013: Journal of Virology
Robert T Gaeta, Rick E Masonbrink, Changzeng Zhao, Abhijit Sanyal, Lakshminarasimhan Krishnaswamy, James A Birchler
Engineered minichromosomes provide efficient platforms for stacking transgenes in crop plants. Methods for modifying these chromosomes in vivo are essential for the development of customizable systems for the removal of selection genes or other sequences and for the addition of new genes. Previous studies have demonstrated that Cre, a site-specific recombinase, could be used to modify lox sites on transgenes on maize minichromosomes; however, these studies demonstrated somatic recombination only, and modified minichromosomes could not be recovered...
June 2013: Chromosoma
Robert Currer, Rachel Van Duyne, Elizabeth Jaworski, Irene Guendel, Gavin Sampey, Ravi Das, Aarthi Narayanan, Fatah Kashanchi
Human T-cell lymphotropic virus type 1 (HTLV-1) has been identified as the causative agent of adult T-cell leukemia (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The virus infects between 15 and 20 million people worldwide of which approximately 2-5% develop ATL. The past 35 years of research have yielded significant insight into the pathogenesis of HTLV-1, including the molecular characterization of Tax, the viral transactivator, and oncoprotein. In spite of these efforts, the mechanisms of oncogenesis of this pleiotropic protein remain to be fully elucidated...
2012: Frontiers in Microbiology
Ludivine Drougat, Stéphanie Olivier-Van Stichelen, Marlène Mortuaire, François Foulquier, Anne-Sophie Lacoste, Jean-Claude Michalski, Tony Lefebvre, Anne-Sophie Vercoutter-Edouart
BACKGROUND: DNA replication represents a critical step of the cell cycle which requires highly controlled and ordered regulatory mechanisms to ensure the integrity of genome duplication. Among a plethora of elements, post-translational modifications (PTMs) ensure the spatiotemporal regulation of pivotal proteins orchestrating cell division. Despite increasing evidences showing that O-GlcNAcylation regulates mitotic events, the impact of this PTM in the early steps of the cell cycle remains poorly understood...
December 2012: Biochimica et Biophysica Acta
Lindsay F Rizzardi, Elizabeth S Dorn, Brian D Strahl, Jeanette Gowen Cook
DNA replication is a highly regulated process that is initiated from replication origins, but the elements of chromatin structure that contribute to origin activity have not been fully elucidated. To identify histone post-translational modifications important for DNA replication, we initiated a genetic screen to identify interactions between genes encoding chromatin-modifying enzymes and those encoding proteins required for origin function in the budding yeast Saccharomyces cerevisiae. We found that enzymes required for histone H3K4 methylation, both the histone methyltransferase Set1 and the E3 ubiquitin ligase Bre1, are required for robust growth of several hypomorphic replication mutants, including cdc6-1...
October 2012: Genetics
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