Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i>
Arabidopsis roots accumulate a complex mixture of dolichols composed of three families, (i.e., short-, medium- and long-chain dolichols), but until now none of the <i>cis</i>-prenyltransferases (CPTs) predicted in the Arabidopsis genome has been considered responsible for their synthesis...
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MDPI AG
2019-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/24/15/2789 |
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| author | Przemyslaw Surowiecki Agnieszka Onysk Katarzyna Manko Ewa Swiezewska Liliana Surmacz |
| author_facet | Przemyslaw Surowiecki Agnieszka Onysk Katarzyna Manko Ewa Swiezewska Liliana Surmacz |
| author_sort | Przemyslaw Surowiecki |
| collection | DOAJ |
| description | Arabidopsis roots accumulate a complex mixture of dolichols composed of three families, (i.e., short-, medium- and long-chain dolichols), but until now none of the <i>cis</i>-prenyltransferases (CPTs) predicted in the Arabidopsis genome has been considered responsible for their synthesis. In this report, using homo- and heterologous (yeast and tobacco) models, we have characterized the <i>AtCPT1</i> gene (At2g23410) which encodes a CPT responsible for the formation of long-chain dolichols, Dol-18 to -23, with Dol-21 dominating, in Arabidopsis. The content of these dolichols was significantly reduced in <i>AtCPT1</i> T-DNA insertion mutant lines and highly increased in <i>AtCPT1</i>-overexpressing plants. Similar to the majority of eukaryotic CPTs, AtCPT1 is localized to the endoplasmic reticulum (ER). Functional complementation tests using yeast <i>rer2</i>Δ or <i>srt1</i>Δ mutants devoid of medium- or long-chain dolichols, respectively, confirmed that this enzyme synthesizes long-chain dolichols, although the dolichol chains thus formed are somewhat shorter than those synthesized in planta. Moreover, AtCPT1 acts as a homomeric CPT and does not need LEW1 for its activity. AtCPT1 is the first plant CPT producing long-chain polyisoprenoids that does not form a complex with the NgBR/NUS1 homologue. |
| first_indexed | 2024-12-13T07:43:28Z |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-13T07:43:28Z |
| publishDate | 2019-07-01 |
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| series | Molecules |
| spelling | doaj.art-b2f7dbbbf0c54f1bad7d873e2edb010a2022-12-21T23:54:55ZengMDPI AGMolecules1420-30492019-07-012415278910.3390/molecules24152789molecules24152789Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i>Przemyslaw Surowiecki0Agnieszka Onysk1Katarzyna Manko2Ewa Swiezewska3Liliana Surmacz4Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, PolandInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, PolandInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, PolandInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, PolandInstitute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, PolandArabidopsis roots accumulate a complex mixture of dolichols composed of three families, (i.e., short-, medium- and long-chain dolichols), but until now none of the <i>cis</i>-prenyltransferases (CPTs) predicted in the Arabidopsis genome has been considered responsible for their synthesis. In this report, using homo- and heterologous (yeast and tobacco) models, we have characterized the <i>AtCPT1</i> gene (At2g23410) which encodes a CPT responsible for the formation of long-chain dolichols, Dol-18 to -23, with Dol-21 dominating, in Arabidopsis. The content of these dolichols was significantly reduced in <i>AtCPT1</i> T-DNA insertion mutant lines and highly increased in <i>AtCPT1</i>-overexpressing plants. Similar to the majority of eukaryotic CPTs, AtCPT1 is localized to the endoplasmic reticulum (ER). Functional complementation tests using yeast <i>rer2</i>Δ or <i>srt1</i>Δ mutants devoid of medium- or long-chain dolichols, respectively, confirmed that this enzyme synthesizes long-chain dolichols, although the dolichol chains thus formed are somewhat shorter than those synthesized in planta. Moreover, AtCPT1 acts as a homomeric CPT and does not need LEW1 for its activity. AtCPT1 is the first plant CPT producing long-chain polyisoprenoids that does not form a complex with the NgBR/NUS1 homologue.https://www.mdpi.com/1420-3049/24/15/2789polyisoprenoidspolyprenolsdolichols<i>cis</i>-prenyltransferaseprotein <i>N</i>-glycosylation<i>Arabidopsis thaliana</i>T-DNA insertion mutant<i>rer2</i>Δ and <i>srt1</i>Δ <i>Saccharomyces cerevisiae</i> mutant |
| spellingShingle | Przemyslaw Surowiecki Agnieszka Onysk Katarzyna Manko Ewa Swiezewska Liliana Surmacz Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> Molecules polyisoprenoids polyprenols dolichols <i>cis</i>-prenyltransferase protein <i>N</i>-glycosylation <i>Arabidopsis thaliana</i> T-DNA insertion mutant <i>rer2</i>Δ and <i>srt1</i>Δ <i>Saccharomyces cerevisiae</i> mutant |
| title | Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> |
| title_full | Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> |
| title_fullStr | Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> |
| title_full_unstemmed | Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> |
| title_short | Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in <i>Arabidopsis thaliana</i> |
| title_sort | long chain polyisoprenoids are synthesized by atcpt1 in i arabidopsis thaliana i |
| topic | polyisoprenoids polyprenols dolichols <i>cis</i>-prenyltransferase protein <i>N</i>-glycosylation <i>Arabidopsis thaliana</i> T-DNA insertion mutant <i>rer2</i>Δ and <i>srt1</i>Δ <i>Saccharomyces cerevisiae</i> mutant |
| url | https://www.mdpi.com/1420-3049/24/15/2789 |
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