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      Terc is dispensable for most of the short-term HPV16 oncogene-mediated phenotypes in mice

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          Abstract

          High-risk human papillomaviruses (HPVs) have been shown in vitro to impinge on telomere homeostasis in a number of ways. However, the in vivo interaction of viruses with the telomere homeostasis apparatus has not been previously explored. Since E6 and E7 are the main viral oncogenes and key for viral replication, we have explored here the short-term phenotypes of the genes in the context of defective telomere homeostasis. We examined the short-term phenotypes of E6 and E7 in a context where the Terc component of the telomerase holoenzyme was knocked out. We determined that Terc was dispensable for most oncogene-mediated phenotypes. Surprisingly, E7-mediated reduction of label retaining cells was found to be in part dependent on the presence of Terc. Under the conditions examined here, there appears to be no compelling evidence Terc is required for most short-term viral oncogene mediated phenotypes. Further studies will elucidate its role in longer-term phenotypes.

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          Most cited references27

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          Telomere shortening and tumor formation by mouse cells lacking telomerase RNA.

          To examine the role of telomerase in normal and neoplastic growth, the telomerase RNA component (mTR) was deleted from the mouse germline. mTR-/- mice lacked detectable telomerase activity yet were viable for the six generations analyzed. Telomerase-deficient cells could be immortalized in culture, transformed by viral oncogenes, and generated tumors in nude mice following transformation. Telomeres were shown to shorten at a rate of 4.8+/-2.4 kb per mTR-/- generation. Cells from the fourth mTR-/- generation onward possessed chromosome ends lacking detectable telomere repeats, aneuploidy, and chromosomal abnormalities, including end-to-end fusions. These results indicate that telomerase is essential for telomere length maintenance but is not required for establishment of cell lines, oncogenic transformation, or tumor formation in mice.
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            Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis.

            Inconsistent with the view that hair follicle stem cells reside in the matrix area of the hair bulb, we found that label-retaining cells exist exclusively in the bulge area of the mouse hair follicle. The bulge consists of a subpopulation of outer root sheath cells located in the midportion of the follicle at the arrector pili muscle attachment site. Keratinocytes in the bulge area are relatively undifferentiated ultrastructurally. They are normally slow cycling, but can be stimulated to proliferate transiently by TPA. Located in a well-protected and nourished environment, these cells mark the lower end of the "permanent" portion of the follicle. Our findings, plus a reevaluation of the literature, suggest that follicular stem cells reside in the bulge region, instead of the lower bulb. This new view provides insights into hair cycle control and the possible involvement of hair follicle stem cells in skin carcinogenesis.
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              Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells.

              Normal human cells undergo a limited number of divisions in culture and enter a non-dividing state called replicative senescence. Senescence is accompanied by several changes, including an increase in inhibitors of cyclin-dependent kinases and telomere shortening. The mechanisms by which viral oncogenes reverse these processes are not fully understood, although a general requirement for oncoproteins such as human papillomavirus E6 and E7 has suggested that the p53 and Rb pathways are targeted. Expression of the catalytic component of telomerase, hTERT, alone significantly extends the lifespan of human fibroblasts. Here we show that telomerase activity is not sufficient for immortalization of human keratinocyte or mammary epithelial cells: we find that neither addition of hTERT nor induction of telomerase activity by E6, both of which are active in maintaining telomere length, results in immortalization. Inactivation of the Rb/p16 pathway by E7 or downregulation of p16 expression, in combination with telomerase activity, however, is able to immortalize epithelial cells efficiently. Elimination of p53 and of the DNA-damage-induced G1 checkpoint is not necessary for immortalization, neither is elimination of p19ARF.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – original draft
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – original draft
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                26 April 2018
                2018
                : 13
                : 4
                : e0196604
                Affiliations
                [001]Department of Biological Sciences, University of Cyprus, Nicosia, Cyprus
                International Centre for Genetic Engineering and Biotechnology, ITALY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-2332-787X
                Article
                PONE-D-18-00803
                10.1371/journal.pone.0196604
                5919663
                29698462
                edf4d179-4fb4-4614-bcb5-a2068010c7d6
                © 2018 Achilleos et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 9 January 2018
                : 16 April 2018
                Page count
                Figures: 5, Tables: 1, Pages: 15
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004117, A.G. Leventis Foundation;
                Award Recipient :
                Funded by: University of Cyprus
                Award Recipient :
                This study was funded by the A.G. Leventis Foundation and the University of Cyprus. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Chromosome Biology
                Chromosomes
                Chromosome Structure and Function
                Telomeres
                Biology and Life Sciences
                Genetics
                Cancer Genetics
                Oncogenes
                Biology and life sciences
                Organisms
                Viruses
                DNA viruses
                Papillomaviruses
                HPV-16
                Biology and Life Sciences
                Microbiology
                Medical Microbiology
                Microbial Pathogens
                Viral Pathogens
                Papillomaviruses
                HPV-16
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Pathogens
                Microbial Pathogens
                Viral Pathogens
                Papillomaviruses
                HPV-16
                Biology and Life Sciences
                Organisms
                Viruses
                Viral Pathogens
                Papillomaviruses
                HPV-16
                Biology and Life Sciences
                Physiology
                Physiological Processes
                Homeostasis
                Medicine and Health Sciences
                Physiology
                Physiological Processes
                Homeostasis
                Medicine and Health Sciences
                Oncology
                Carcinogenesis
                Biology and Life Sciences
                Anatomy
                Integumentary System
                Hair
                Hair Follicles
                Medicine and Health Sciences
                Anatomy
                Integumentary System
                Hair
                Hair Follicles
                Biology and Life Sciences
                Anatomy
                Integumentary System
                Skin
                Hair Follicles
                Medicine and Health Sciences
                Anatomy
                Integumentary System
                Skin
                Hair Follicles
                Medicine and Health Sciences
                Oncology
                Cancer Risk Factors
                Viruses and Cancer
                Biology and Life Sciences
                Microbiology
                Virology
                Viruses and Cancer
                Biology and Life Sciences
                Anatomy
                Biological Tissue
                Epithelium
                Medicine and Health Sciences
                Anatomy
                Biological Tissue
                Epithelium
                Custom metadata
                All relevant data are available from Figshare at https://figshare.com/articles/The_effect_of_HPV16_in_the_absence_of_Terc/6120533.

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