Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope
In all eukaryotic cells, the nucleus forms a prominent cellular compartment containing the cell’s nuclear genome. Although structurally similar, animal and plant nuclei differ substantially in details of their architecture. One example is the nuclear lamina, a layer of tightly interconnected filamen...
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MDPI AG
2016-03-01
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| Series: | Viruses |
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| Online Access: | http://www.mdpi.com/1999-4915/8/3/73 |
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| author | Christian E. Lamm Katrin Link Sabrina Wagner Jens Milbradt Manfred Marschall Uwe Sonnewald |
| author_facet | Christian E. Lamm Katrin Link Sabrina Wagner Jens Milbradt Manfred Marschall Uwe Sonnewald |
| author_sort | Christian E. Lamm |
| collection | DOAJ |
| description | In all eukaryotic cells, the nucleus forms a prominent cellular compartment containing the cell’s nuclear genome. Although structurally similar, animal and plant nuclei differ substantially in details of their architecture. One example is the nuclear lamina, a layer of tightly interconnected filament proteins (lamins) underlying the nuclear envelope of metazoans. So far no orthologous lamin genes could be detected in plant genomes and putative lamin-like proteins are only poorly described in plants. To probe for potentially conserved features of metazoan and plant nuclear envelopes, we ectopically expressed the core nuclear egress proteins of human cytomegalovirus pUL50 and pUL53 in plant cells. pUL50 localizes to the inner envelope of metazoan nuclei and recruits the nuclear localized pUL53 to it, forming heterodimers. Upon expression in plant cells, a very similar localization pattern of both proteins could be determined. Notably, pUL50 is specifically targeted to the plant nuclear envelope in a rim-like fashion, a location to which coexpressed pUL53 becomes strictly corecruited from its initial nucleoplasmic distribution. Using pUL50 as bait in a yeast two-hybrid screening, the cytoplasmic re-initiation supporting protein RISP could be identified. Interaction of pUL50 and RISP could be confirmed by coexpression and coimmunoprecipitation in mammalian cells and by confocal laser scanning microscopy in plant cells, demonstrating partial pUL50-RISP colocalization in areas of the nuclear rim and other intracellular compartments. Thus, our study provides strong evidence for conserved structural features of plant and metazoan nuclear envelops and identifies RISP as a potential pUL50-interacting plant protein. |
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| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-10T10:54:00Z |
| publishDate | 2016-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-ae969932047145649c40b065863b98732022-12-22T01:51:56ZengMDPI AGViruses1999-49152016-03-01837310.3390/v8030073v8030073Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear EnvelopeChristian E. Lamm0Katrin Link1Sabrina Wagner2Jens Milbradt3Manfred Marschall4Uwe Sonnewald5Division of Biochemistry, Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, Erlangen 91058, GermanyDivision of Biochemistry, Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, Erlangen 91058, GermanyInstitute for Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nuremberg, Schloßgarten 4, Erlangen 91054, GermanyInstitute for Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nuremberg, Schloßgarten 4, Erlangen 91054, GermanyInstitute for Clinical and Molecular Virology, Friedrich-Alexander University Erlangen-Nuremberg, Schloßgarten 4, Erlangen 91054, GermanyDivision of Biochemistry, Department of Biology, Friedrich-Alexander University Erlangen-Nuremberg, Staudtstrasse 5, Erlangen 91058, GermanyIn all eukaryotic cells, the nucleus forms a prominent cellular compartment containing the cell’s nuclear genome. Although structurally similar, animal and plant nuclei differ substantially in details of their architecture. One example is the nuclear lamina, a layer of tightly interconnected filament proteins (lamins) underlying the nuclear envelope of metazoans. So far no orthologous lamin genes could be detected in plant genomes and putative lamin-like proteins are only poorly described in plants. To probe for potentially conserved features of metazoan and plant nuclear envelopes, we ectopically expressed the core nuclear egress proteins of human cytomegalovirus pUL50 and pUL53 in plant cells. pUL50 localizes to the inner envelope of metazoan nuclei and recruits the nuclear localized pUL53 to it, forming heterodimers. Upon expression in plant cells, a very similar localization pattern of both proteins could be determined. Notably, pUL50 is specifically targeted to the plant nuclear envelope in a rim-like fashion, a location to which coexpressed pUL53 becomes strictly corecruited from its initial nucleoplasmic distribution. Using pUL50 as bait in a yeast two-hybrid screening, the cytoplasmic re-initiation supporting protein RISP could be identified. Interaction of pUL50 and RISP could be confirmed by coexpression and coimmunoprecipitation in mammalian cells and by confocal laser scanning microscopy in plant cells, demonstrating partial pUL50-RISP colocalization in areas of the nuclear rim and other intracellular compartments. Thus, our study provides strong evidence for conserved structural features of plant and metazoan nuclear envelops and identifies RISP as a potential pUL50-interacting plant protein.http://www.mdpi.com/1999-4915/8/3/73human cytomegaloviruspUL50pUL53nuclear envelopeplant cellsre-initiation supporting protein RISP |
| spellingShingle | Christian E. Lamm Katrin Link Sabrina Wagner Jens Milbradt Manfred Marschall Uwe Sonnewald Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope Viruses human cytomegalovirus pUL50 pUL53 nuclear envelope plant cells re-initiation supporting protein RISP |
| title | Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope |
| title_full | Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope |
| title_fullStr | Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope |
| title_full_unstemmed | Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope |
| title_short | Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope |
| title_sort | human cytomegalovirus nuclear egress proteins ectopically expressed in the heterologous environment of plant cells are strictly targeted to the nuclear envelope |
| topic | human cytomegalovirus pUL50 pUL53 nuclear envelope plant cells re-initiation supporting protein RISP |
| url | http://www.mdpi.com/1999-4915/8/3/73 |
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