Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens
Abstract
Results from live cell imaging of fluorescently tagged Cellulose Synthase (CESA) proteins in Cellulose Synthesis Complexes (CSCs) have enhanced our understanding of cellulose biosynthesis, including the mechanisms of action of cellulose synthesis inhibitors. However, this method has been applied only in Arabidopsis thaliana and Brachypodium distachyon thus far. Results from freeze fracture electron microscopy of protonemal filaments of the moss Funaria hygrometrica indicate that a cellulose synthesis inhibitor, 2,6-dichlorobenzonitrile (DCB), fragments CSCs and clears them from the plasma membrane. This differs from Arabidopsis, in which DCB causes CSC accumulation in the plasma membrane and a different cellulose synthesis inhibitor, isoxaben, clears CSCs from the plasma membrane. In this study, live cell imaging of the moss Physcomitrella patens indicated that DCB and isoxaben have little effect on protonemal growth rates, and that only DCB causes tip rupture. Live cell imaging of mEGFP-PpCESA5 and mEGFP-PpCESA8 showed that DCB and isoxaben substantially reduced CSC movement, but had no measureable effect on CSC density in the plasma membrane. These results suggest that DCB and isoxaben have similar effects on CSC movement in P. patens and Arabidopsis, but have different effects on CSC intracellular trafficking, cell growth and cell integrity in these divergent plant lineages.
- Authors:
-
[2];
[2];
[1]
- Univ. of Rhode Island, Kingston, RI (United States)
- Pennsylvania State Univ., University Park, PA (United States)
- Worcester Polytechnic Inst., Worcester, MA (United States)
- Univ. of Massachusetts, Amherst, MA (United States)
- Publication Date:
- Research Org.:
- Pennsylvania State Univ., University Park, PA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1500047
- Grant/Contract Number:
- SC0001090
- Resource Type:
- Journal Article: Accepted Manuscript
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Tran, Mai L., McCarthy, Thomas W., Sun, Hao, Wu, Shu-Zon, Norris, Joanna H., Bezanilla, Magdalena, Vidali, Luis, Anderson, Charles T., and Roberts, Alison W. Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens. United States: N. p., 2018.
Web. doi:10.1038/s41598-017-18994-4.
Tran, Mai L., McCarthy, Thomas W., Sun, Hao, Wu, Shu-Zon, Norris, Joanna H., Bezanilla, Magdalena, Vidali, Luis, Anderson, Charles T., & Roberts, Alison W. Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens. United States. doi:10.1038/s41598-017-18994-4.
Tran, Mai L., McCarthy, Thomas W., Sun, Hao, Wu, Shu-Zon, Norris, Joanna H., Bezanilla, Magdalena, Vidali, Luis, Anderson, Charles T., and Roberts, Alison W. Mon .
"Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens". United States. doi:10.1038/s41598-017-18994-4. https://www.osti.gov/servlets/purl/1500047.
@article{osti_1500047,
title = {Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens},
author = {Tran, Mai L. and McCarthy, Thomas W. and Sun, Hao and Wu, Shu-Zon and Norris, Joanna H. and Bezanilla, Magdalena and Vidali, Luis and Anderson, Charles T. and Roberts, Alison W.},
abstractNote = {Results from live cell imaging of fluorescently tagged Cellulose Synthase (CESA) proteins in Cellulose Synthesis Complexes (CSCs) have enhanced our understanding of cellulose biosynthesis, including the mechanisms of action of cellulose synthesis inhibitors. However, this method has been applied only in Arabidopsis thaliana and Brachypodium distachyon thus far. Results from freeze fracture electron microscopy of protonemal filaments of the moss Funaria hygrometrica indicate that a cellulose synthesis inhibitor, 2,6-dichlorobenzonitrile (DCB), fragments CSCs and clears them from the plasma membrane. This differs from Arabidopsis, in which DCB causes CSC accumulation in the plasma membrane and a different cellulose synthesis inhibitor, isoxaben, clears CSCs from the plasma membrane. In this study, live cell imaging of the moss Physcomitrella patens indicated that DCB and isoxaben have little effect on protonemal growth rates, and that only DCB causes tip rupture. Live cell imaging of mEGFP-PpCESA5 and mEGFP-PpCESA8 showed that DCB and isoxaben substantially reduced CSC movement, but had no measureable effect on CSC density in the plasma membrane. These results suggest that DCB and isoxaben have similar effects on CSC movement in P. patens and Arabidopsis, but have different effects on CSC intracellular trafficking, cell growth and cell integrity in these divergent plant lineages.},
doi = {10.1038/s41598-017-18994-4},
journal = {Scientific Reports},
issn = {2045-2322},
number = 1,
volume = 8,
place = {United States},
year = {2018},
month = {1}
}
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Figure 1: Expression of mEGFP-PpCESAs rescue the cognate mutant phenotypes. (a) Transformation of the gametophore defective ppcesa5KO-2 line with Ubi::GFP-PpCESA5 restores wild type gametophore development. (b) Transformation of the ppcesa8KO-5B line, which is characterized by weak midrib birefringence due to reduced secondary cell wall deposition, was partially restored to themore »
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