The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae
Agrobacterium tumefaciens is capable of gene transfer to both plant and non-plant organisms. Indeed, upon infection of eukaryotic cells, Agrobacterium tumefaciens transfers a piece of its tumor inducing (Ti)-plasmid, called T-DNA, to the host cell nucleus, which subsequently integrates into the host...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Ferdowsi University of Mashhad
2013-12-01
|
| Series: | Journal of Cell and Molecular Research |
| Subjects: | |
| Online Access: | https://jcmr.um.ac.ir/article_27024_e466dfb8ad62888472b5e7d58970f2c9.pdf |
| _version_ | 1818976013036552192 |
|---|---|
| author | Jalal Soltani Jonathan A. Lal G. Paul H. van Heusden Paul J.J. Hooykaas |
| author_facet | Jalal Soltani Jonathan A. Lal G. Paul H. van Heusden Paul J.J. Hooykaas |
| author_sort | Jalal Soltani |
| collection | DOAJ |
| description | Agrobacterium tumefaciens is capable of gene transfer to both plant and non-plant organisms. Indeed, upon infection of eukaryotic cells, Agrobacterium tumefaciens transfers a piece of its tumor inducing (Ti)-plasmid, called T-DNA, to the host cell nucleus, which subsequently integrates into the host genome. The VirD2 virulence protein which has relaxase endonuclease activities covalently binds to the 5'end of T-DNA and facilitates its transfer, nuclear localization and integration into the host genome in collaboration with the interacting proteins of the host cell. The VirD2 is essential for Agrobacterium–mediated transformation of both plants and non-plant cells. Here, using yeast Green Flourescent Protein (yGFP) technology, we studied the subcellular localization of VirD2, expressed in the model eukaryote Saccharomyces cerevisiae. Fluorescence microscopy showed that an N-terminal yGFP fusion of VirD2 (i.e. 5' GFP-VirD2 3'), was located in the nucleus of yeast. With C-terminal fusions of VirD2 to yGFP (i.e. 5' VirD2-GFP 3'), no particular subcellular concentration of fluorescence was seen. This further confirms nuclear localization of VirD2 in eukaryotic cells and more importantly highlights the role of Nuclear Localization Signal sequences (NLS) of the C-terminal of VirD2 in this phenomenon. |
| first_indexed | 2024-12-20T16:05:05Z |
| format | Article |
| id | doaj.art-55888adf581b42deb3bab3404d92aec1 |
| institution | Directory Open Access Journal |
| issn | 2008-9147 2717-3364 |
| language | English |
| last_indexed | 2024-12-20T16:05:05Z |
| publishDate | 2013-12-01 |
| publisher | Ferdowsi University of Mashhad |
| record_format | Article |
| series | Journal of Cell and Molecular Research |
| spelling | doaj.art-55888adf581b42deb3bab3404d92aec12022-12-21T19:34:10ZengFerdowsi University of MashhadJournal of Cell and Molecular Research2008-91472717-33642013-12-0152606410.22067/jcmr.v5i2.1354127024The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiaeJalal SoltaniJonathan A. LalG. Paul H. van HeusdenPaul J.J. HooykaasAgrobacterium tumefaciens is capable of gene transfer to both plant and non-plant organisms. Indeed, upon infection of eukaryotic cells, Agrobacterium tumefaciens transfers a piece of its tumor inducing (Ti)-plasmid, called T-DNA, to the host cell nucleus, which subsequently integrates into the host genome. The VirD2 virulence protein which has relaxase endonuclease activities covalently binds to the 5'end of T-DNA and facilitates its transfer, nuclear localization and integration into the host genome in collaboration with the interacting proteins of the host cell. The VirD2 is essential for Agrobacterium–mediated transformation of both plants and non-plant cells. Here, using yeast Green Flourescent Protein (yGFP) technology, we studied the subcellular localization of VirD2, expressed in the model eukaryote Saccharomyces cerevisiae. Fluorescence microscopy showed that an N-terminal yGFP fusion of VirD2 (i.e. 5' GFP-VirD2 3'), was located in the nucleus of yeast. With C-terminal fusions of VirD2 to yGFP (i.e. 5' VirD2-GFP 3'), no particular subcellular concentration of fluorescence was seen. This further confirms nuclear localization of VirD2 in eukaryotic cells and more importantly highlights the role of Nuclear Localization Signal sequences (NLS) of the C-terminal of VirD2 in this phenomenon.https://jcmr.um.ac.ir/article_27024_e466dfb8ad62888472b5e7d58970f2c9.pdfvird2nuclear deliverygfpagrobacteriumsaccharomyces cerevisiae |
| spellingShingle | Jalal Soltani Jonathan A. Lal G. Paul H. van Heusden Paul J.J. Hooykaas The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae Journal of Cell and Molecular Research vird2 nuclear delivery gfp agrobacterium saccharomyces cerevisiae |
| title | The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae |
| title_full | The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae |
| title_fullStr | The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae |
| title_full_unstemmed | The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae |
| title_short | The significance of C-terminal NLS sequences of VirD2 in its nuclear localization in Saccharomyces cerevisiae |
| title_sort | significance of c terminal nls sequences of vird2 in its nuclear localization in saccharomyces cerevisiae |
| topic | vird2 nuclear delivery gfp agrobacterium saccharomyces cerevisiae |
| url | https://jcmr.um.ac.ir/article_27024_e466dfb8ad62888472b5e7d58970f2c9.pdf |
| work_keys_str_mv | AT jalalsoltani thesignificanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT jonathanalal thesignificanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT gpaulhvanheusden thesignificanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT pauljjhooykaas thesignificanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT jalalsoltani significanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT jonathanalal significanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT gpaulhvanheusden significanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae AT pauljjhooykaas significanceofcterminalnlssequencesofvird2initsnuclearlocalizationinsaccharomycescerevisiae |