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      The exon junction complex factor Y14 is dynamic in the nucleus of the beetle Tribolium castaneum during late oogenesis


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          The oocyte chromosomes of the red flour beetle, Tribolium castaneum, are gathered into a knot, forming a karyosphere at the diplotene stage of meiotic prophase. Chromatin rearrangement, which is a characteristic feature of oocyte maturation, is well documented. The T. castaneum karyosphere is surrounded by a complex extrachromosomal structure termed the karyosphere capsule. The capsule contains the vast majority of oocyte RNA. We have previously shown using a BrUTP assay that oocyte chromosomes in T. castaneum maintain residual transcription up to the very end of oocyte maturation. Karyosphere transcription requires evidently not only transcription factors but also mRNA processing factors, including the components of the exon junction complex with its core component, the splicing factor Y14. We employed a gene engineering approach with injection of mRNA derived from the Myc-tagged Y14 plasmid-based construct in order to monitor the newly synthesized fusion protein in the oocyte nuclei.


          Our preliminary data have been presented as a brief correspondence elsewhere. Here, we provide a full-length article including immunoelectron-microscopy localization data on Y14–Myc distribution in the nucleus of previtellogenic and vitellogenic oocytes. The injections of the fusion protein Y14–Myc mRNA into the oocytes showed a dynamic pattern of the protein distribution. At the previtellogenic stage, there are two main locations for the protein: SC35 domains (the analogues of interchromatin granule clusters or nuclear speckles) and the karyosphere capsule. At the vitellogenic stage, SC35 domains were devoid of labels, and Y14–Myc was found in the perichromatin region of the karyosphere, presumably at the places of residual transcription. We show that karyosphere formation is accompanied by the movement of a nuclear protein while the residual transcription occurs during genome inactivation.


          Our data indicate that the karyosphere capsule, being a destination site for a protein involved in mRNA splicing and export, is not only a specializes part of nuclear matrix separating the karyosphere from the products of chromosome activity, as believed previously, but represents a special nuclear compartment involved in the processes of gene expression in the case the karyosphere retains residual transcription activity.

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          The online version of this article (10.1186/s13039-017-0342-4) contains supplementary material, which is available to authorized users.

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          The genome of the model beetle and pest Tribolium castaneum.

          Tribolium castaneum is a member of the most species-rich eukaryotic order, a powerful model organism for the study of generalized insect development, and an important pest of stored agricultural products. We describe its genome sequence here. This omnivorous beetle has evolved the ability to interact with a diverse chemical environment, as shown by large expansions in odorant and gustatory receptors, as well as P450 and other detoxification enzymes. Development in Tribolium is more representative of other insects than is Drosophila, a fact reflected in gene content and function. For example, Tribolium has retained more ancestral genes involved in cell-cell communication than Drosophila, some being expressed in the growth zone crucial for axial elongation in short-germ development. Systemic RNA interference in T. castaneum functions differently from that in Caenorhabditis elegans, but nevertheless offers similar power for the elucidation of gene function and identification of targets for selective insect control.
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            Characteristics of a human cell line transformed by DNA from human adenovirus type 5.

            Human embryonic kidney cells have been transformed by exposing cells to sheared fragments of adenovirus type 5 DNA. The transformed cells (designated 293 cells) exhibited many of the characteristics of transformation including the elaboration of a virus-specific tumour antigen. Analysis of the polypeptides synthesized in the 293 cells by labelling with 35S-methionine and SDS PAGE showed a variable pattern of synthesis, different in a number of respects from that seen in otheruman cells. On labelling the surface of cells by lactoperoxidase catalysed radio-iodination, the absence of a labelled polypeptide analogous to the 250 K (LETS) glycoprotein was noted. Hybridization of labelled cellular RNA with restriction fragments of adenovirus type 5 DNA indicated transcription of a portion of the adenovirus genome at the conventional left hand end.
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              Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product.

              Six monoclonal antibodies have been isolated from mice immunized with synthetic peptide immunogens whose sequences are derived from that of the human c-myc gene product. Five of these antibodies precipitate p62c-myc from human cells, and three of these five also recognize the mouse c-myc gene product. None of the antibodies sees the chicken p110gag-myc protein. All six antibodies recognize immunoblotted p62c-myc. These reagents also provide the basis for an immunoblotting assay by which to quantitate p62c-myc in cells.

                Author and article information

                dbogol@mail.ru , dmitr@incras.ru
                opodg@yahoo.com , o.podgornaya@spbu.ru
                Mol Cytogenet
                Mol Cytogenet
                Molecular Cytogenetics
                BioMed Central (London )
                9 November 2017
                9 November 2017
                : 10
                : 41
                [1 ]ISNI 0000 0000 9629 3848, GRID grid.418947.7, Laboratory of Cell Morphology, Institute of Cytology, Russian Academy of Sciences, ; St. Petersburg, 194064 Russia
                [2 ]Federal Almazov North-West Medical Research Centre, St. Petersburg, 197341 Russia
                [3 ]ISNI 0000 0001 0413 4629, GRID grid.35915.3b, ITMO University, Institute of Translational Medicine, ; St. Petersburg, 197101 Russia
                [4 ]ISNI 0000 0001 2289 6897, GRID grid.15447.33, Department of Cytology and Histology, Faculty of Biology, , St. Petersburg State University, ; St. Petersburg, 199034 Russia
                [5 ]ISNI 0000 0004 0637 7917, GRID grid.440624.0, Far Eastern Federal University, School of Biomedicine, ; Vladivostok, 690950 Russia
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                : 8 May 2017
                : 27 October 2017
                Funded by: Government of Russian Federation
                Award ID: 074-U01
                Award Recipient :
                Funded by: The granting program ‘Molecular and cell biology’ of the Russian Academy of Sciences
                Award ID: 01201457147
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100006769, Russian Science Foundation;
                Award ID: 15-15-20026
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100002261, Russian Foundation for Basic Research;
                Award ID: 15-04-01857
                Award Recipient :
                Custom metadata
                © The Author(s) 2017

                tribolium castaneum,y14 (tsunagi),oocyte nucleus,karyosphere,karyosphere capsule,interchromatin granule clusters,microinjections,molecular cloning,gene manipulation,transcription in vitro


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