Cloning and molecular characterisation of a Δ8-sphingolipid-desaturase from Nicotiana tabacum closely related to Δ6-acyl-desaturases

Cloning and molecular characterisation of a Δ8-sphingolipid-desaturase from Nicotiana tabacum... Investigation on the absence of Δ6-desaturase activity in Nicotiana tabacum has led to the cloning of a new desaturase gene from this organism (NTDXDES) that exhibited unexpected biochemical activity. Cladistic analysis shows clustering of NTDXDES together with functional Δ6-acyl-desaturases of near Solanales plants, such as Borago and Echium. This group lies apart from that of previously characterised Δ8-sphingolipid-desaturases, which also includes two putative tobacco members identified in this study. Moreover, strong expression of NTDXDES is found in leaves, flowers, fruits and developing seeds of tobacco plants that is highly dependent on the development phase, with transcriptional activity being higher at stages of active tissue growth. This pattern is similar to that showed by Δ6-acyl-desaturases characterised in Boraginaceae species. However, functional assays using a yeast expression system revealed that the protein encoded by NTDXDES lacks Δ6-desaturase activity, but instead it is able to desaturate sphingolipid substrates by introducing a double bond on the Δ8-position. These data indicate that NTDXDES represent a novel desaturase gene placed in a different evolutionary lineage to that of previously characterised Δ8-desaturases. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Cloning and molecular characterisation of a Δ8-sphingolipid-desaturase from Nicotiana tabacum closely related to Δ6-acyl-desaturases

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 2007 by Springer Science+Business Media B.V.
Subject
Life Sciences; Plant Pathology; Biochemistry, general; Plant Sciences
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1007/s11103-007-9148-9
Publisher site
See Article on Publisher Site

Abstract

Investigation on the absence of Δ6-desaturase activity in Nicotiana tabacum has led to the cloning of a new desaturase gene from this organism (NTDXDES) that exhibited unexpected biochemical activity. Cladistic analysis shows clustering of NTDXDES together with functional Δ6-acyl-desaturases of near Solanales plants, such as Borago and Echium. This group lies apart from that of previously characterised Δ8-sphingolipid-desaturases, which also includes two putative tobacco members identified in this study. Moreover, strong expression of NTDXDES is found in leaves, flowers, fruits and developing seeds of tobacco plants that is highly dependent on the development phase, with transcriptional activity being higher at stages of active tissue growth. This pattern is similar to that showed by Δ6-acyl-desaturases characterised in Boraginaceae species. However, functional assays using a yeast expression system revealed that the protein encoded by NTDXDES lacks Δ6-desaturase activity, but instead it is able to desaturate sphingolipid substrates by introducing a double bond on the Δ8-position. These data indicate that NTDXDES represent a novel desaturase gene placed in a different evolutionary lineage to that of previously characterised Δ8-desaturases.

Journal

Plant Molecular BiologySpringer Journals

Published: Feb 27, 2007

References

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