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An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus
The highly variable cytoplasmic N-terminus of the plant diacylglycerol acyltransferase 1 (DGAT1) has been shown to have roles in oligomerization as well as allostery; however, the biological significance of the variation within this region is not understood. Comparing the coding sequences over the v...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9523541/ https://www.ncbi.nlm.nih.gov/pubmed/36186081 http://dx.doi.org/10.3389/fpls.2022.951389 |
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author | Winichayakul, Somrutai Curran, Amy Moraga, Roger Cookson, Ruth Xue, Hong Crowther, Tracey Roldan, Marissa Bryan, Greg Roberts, Nick |
author_facet | Winichayakul, Somrutai Curran, Amy Moraga, Roger Cookson, Ruth Xue, Hong Crowther, Tracey Roldan, Marissa Bryan, Greg Roberts, Nick |
author_sort | Winichayakul, Somrutai |
collection | PubMed |
description | The highly variable cytoplasmic N-terminus of the plant diacylglycerol acyltransferase 1 (DGAT1) has been shown to have roles in oligomerization as well as allostery; however, the biological significance of the variation within this region is not understood. Comparing the coding sequences over the variable N-termini revealed the Poaceae DGAT1s contain relatively high GC compositional gradients as well as numerous direct and inverted repeats in this region. Using a variety of reciprocal chimeric DGAT1s from angiosperms we show that related N-termini had similar effects (positive or negative) on the accumulation of the recombinant protein in Saccharomyces cerevisiae. When expressed in Camelina sativa seeds the recombinant proteins of specific chimeras elevated total lipid content of the seeds as well as increased seed size. In addition, we combine N- and C-terminal as well as internal tags with high pH membrane reformation, protease protection and differential permeabilization. This led us to conclude the C-terminus is in the ER lumen; this contradicts earlier reports of the cytoplasmic location of plant DGAT1 C-termini. |
format | Online Article Text |
id | pubmed-9523541 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-95235412022-10-01 An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus Winichayakul, Somrutai Curran, Amy Moraga, Roger Cookson, Ruth Xue, Hong Crowther, Tracey Roldan, Marissa Bryan, Greg Roberts, Nick Front Plant Sci Plant Science The highly variable cytoplasmic N-terminus of the plant diacylglycerol acyltransferase 1 (DGAT1) has been shown to have roles in oligomerization as well as allostery; however, the biological significance of the variation within this region is not understood. Comparing the coding sequences over the variable N-termini revealed the Poaceae DGAT1s contain relatively high GC compositional gradients as well as numerous direct and inverted repeats in this region. Using a variety of reciprocal chimeric DGAT1s from angiosperms we show that related N-termini had similar effects (positive or negative) on the accumulation of the recombinant protein in Saccharomyces cerevisiae. When expressed in Camelina sativa seeds the recombinant proteins of specific chimeras elevated total lipid content of the seeds as well as increased seed size. In addition, we combine N- and C-terminal as well as internal tags with high pH membrane reformation, protease protection and differential permeabilization. This led us to conclude the C-terminus is in the ER lumen; this contradicts earlier reports of the cytoplasmic location of plant DGAT1 C-termini. Frontiers Media S.A. 2022-09-16 /pmc/articles/PMC9523541/ /pubmed/36186081 http://dx.doi.org/10.3389/fpls.2022.951389 Text en Copyright © 2022 Winichayakul, Curran, Moraga, Cookson, Xue, Crowther, Roldan, Bryan and Roberts. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Winichayakul, Somrutai Curran, Amy Moraga, Roger Cookson, Ruth Xue, Hong Crowther, Tracey Roldan, Marissa Bryan, Greg Roberts, Nick An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title | An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title_full | An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title_fullStr | An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title_full_unstemmed | An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title_short | An alternative angiosperm DGAT1 topology and potential motifs in the N-terminus |
title_sort | alternative angiosperm dgat1 topology and potential motifs in the n-terminus |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9523541/ https://www.ncbi.nlm.nih.gov/pubmed/36186081 http://dx.doi.org/10.3389/fpls.2022.951389 |
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