CMV-promoter driven codon-optimized expression alters the assembly type and morphology of a reconstituted HERV-K(HML-2)
The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have nevertheless been subjected to mutations or deletions that render expressed particles non-infectious. Moreover, these post-insertional mutatio...
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Published in | Viruses Vol. 6; no. 11; pp. 4332 - 4345 |
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Abstract | The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have nevertheless been subjected to mutations or deletions that render expressed particles non-infectious. Moreover, these post-insertional mutations hamper the analysis of the general biological properties of this ancient virus family. The expression of consensus sequences and sequences of elements with reverted post-insertional mutations has therefore been very instrumental in overcoming this limitation. We investigated the particle morphology of a recently reconstituted HERV-K113 element termed oriHERV-K113 using thin-section electron microscopy (EM) and could demonstrate that strong overexpression by substitution of the 5'LTR for a CMV promoter and partial codon optimization altered the virus assembly type and morphology. This included a conversion from the regular C-type to an A-type morphology with a mass of cytoplasmic immature cores tethered to the cell membrane and the membranes of vesicles. Overexpression permitted the release and maturation of virions but reduced the envelope content. A weaker boost of virus expression by Staufen-1 was not sufficient to induce these morphological alterations. |
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AbstractList | The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have nevertheless been subjected to mutations or deletions that render expressed particles non-infectious. Moreover, these post-insertional mutations hamper the analysis of the general biological properties of this ancient virus family. The expression of consensus sequences and sequences of elements with reverted post-insertional mutations has therefore been very instrumental in overcoming this limitation. We investigated the particle morphology of a recently reconstituted HERV-K113 element termed oriHERV-K113 using thin-section electron microscopy (EM) and could demonstrate that strong overexpression by substitution of the 5'LTR for a CMV promoter and partial codon optimization altered the virus assembly type and morphology. This included a conversion from the regular C-type to an A-type morphology with a mass of cytoplasmic immature cores tethered to the cell membrane and the membranes of vesicles. Overexpression permitted the release and maturation of virions but reduced the envelope content. A weaker boost of virus expression by Staufen-1 was not sufficient to induce these morphological alterations. |
Author | Bannert, Norbert Hohn, Oliver Mostafa, Saeed Hanke, Kirsten Lausch, Veronika Zimmermann, Anja |
AuthorAffiliation | HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany; E-Mails: HohnO@rki.de (O.H.); HankeK@rki.de (K.H.); LauschV@rki.de (V.L.); ZimmermannA@rki.de (A.Z.); MostafaS@rki.de (S.M.) |
AuthorAffiliation_xml | – name: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany; E-Mails: HohnO@rki.de (O.H.); HankeK@rki.de (K.H.); LauschV@rki.de (V.L.); ZimmermannA@rki.de (A.Z.); MostafaS@rki.de (S.M.) |
Author_xml | – sequence: 1 givenname: Oliver surname: Hohn fullname: Hohn, Oliver email: HohnO@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. HohnO@rki.de – sequence: 2 givenname: Kirsten surname: Hanke fullname: Hanke, Kirsten email: HankeK@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. HankeK@rki.de – sequence: 3 givenname: Veronika surname: Lausch fullname: Lausch, Veronika email: LauschV@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. LauschV@rki.de – sequence: 4 givenname: Anja surname: Zimmermann fullname: Zimmermann, Anja email: ZimmermannA@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. ZimmermannA@rki.de – sequence: 5 givenname: Saeed surname: Mostafa fullname: Mostafa, Saeed email: MostafaS@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. MostafaS@rki.de – sequence: 6 givenname: Norbert surname: Bannert fullname: Bannert, Norbert email: BannertN@rki.de organization: HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. BannertN@rki.de |
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Cites_doi | 10.1089/104303400750038507 10.1128/JVI.75.22.10991-11001.2001 10.1126/science.1235148 10.1186/1742-4690-8-90 10.1099/0022-1317-82-3-591 10.1016/S0065-230X(08)60223-7 10.1128/JVI.03031-12 10.1038/35001608 10.1128/JVI.01368-09 10.1099/0022-1317-64-12-2549 10.1099/0022-1317-80-1-255 10.1101/gr.5565706 10.1016/j.immuni.2012.07.018 10.1038/nature11599 10.1371/journal.ppat.0030010 10.1158/0008-5472.CAN-04-2983 10.1146/annurev.genom.7.080505.115700 10.1002/ajpa.20459 10.1186/1742-4690-8-30 10.1016/S0960-9822(01)00455-9 10.1186/1742-4690-10-140 10.1016/S0960-9822(01)00553-X |
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Snippet | The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have... |
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StartPage | 4332 |
SubjectTerms | A-type particles budding Cell Line Chromosomes Cloning Cytomegalovirus Cytomegalovirus - genetics endogenous retrovirus Endogenous Retroviruses - genetics Endogenous Retroviruses - physiology Endogenous Retroviruses - ultrastructure Gene Expression Regulation, Viral Genomes HERV-K(HML-2) HERV-K113 Humans Microscopy Microscopy, Electron, Transmission Morphology Mutation Plasmids Promoter Regions, Genetic Protein expression Virion - ultrastructure Virus Assembly Viruses |
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Title | CMV-promoter driven codon-optimized expression alters the assembly type and morphology of a reconstituted HERV-K(HML-2) |
URI | https://www.ncbi.nlm.nih.gov/pubmed/25393897 https://www.proquest.com/docview/1635733663/abstract/ https://search.proquest.com/docview/1625347202 https://search.proquest.com/docview/1642611214 https://pubmed.ncbi.nlm.nih.gov/PMC4246225 https://doaj.org/article/5ed61a8a2029445a966bc62cd1ac747a |
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