skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1{sup A903V} and CESA3{sup T942I} of cellulose synthase

Abstract

The mechanisms underlying the biosynthesis of cellulose in plants are complex and still poorly understood. A central question concerns the mechanism of microfibril structure and how this is linked to the catalytic polymerization action of cellulose synthase (CESA). Furthermore, it remains unclear whether modification of cellulose microfibril structure can be achieved genetically, which could be transformative in a bio-based economy. To explore these processes in planta, we developed a chemical genetic toolbox of pharmacological inhibitors and corresponding resistance-conferring point mutations in the C-terminal transmembrane domain region of CESA1{sup A903V} and CESA3{sup T942I} in Arabidopsis thaliana. Using {sup 13}C solid-state nuclear magnetic resonance spectroscopy and X-ray diffraction, we show that the cellulose microfibrils displayed reduced width and an additional cellulose C4 peak indicative of a degree of crystallinity that is intermediate between the surface and interior glucans of wild type, suggesting a difference in glucan chain association during microfibril formation. Consistent with measurements of lower microfibril crystallinity, cellulose extracts from mutated CESA1{sup A903V} and CESA3{sup T942I} displayed greater saccharification efficiency than wild type. Using live-cell imaging to track fluorescently labeled CESA, we found that these mutants show increased CESA velocities in the plasma membrane, an indication of increased polymerization rate. Collectively,more » these data suggest that CESA1{sup A903V} and CESA3{sup T942I} have modified microfibril structure in terms of crystallinity and suggest that in plants, as in bacteria, crystallization biophysically limits polymerization.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Ames Laboratory (AMES), Ames, IA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1044640
Report Number(s):
IS-J 7666
DOE Contract Number:  
DE-AC02-07CH11358
Resource Type:
Journal Article
Journal Name:
Proceedings of the National Academy of Sciences
Additional Journal Information:
Journal Volume: 2012; Journal Issue: February
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; cell wall polysaccharide quinoxyphen

Citation Formats

Harris, Darby, Corbin, Kendall, Wang, Tuo, Gutierrez, Ryan, Bertolo, Ana, Petti, Caroalberto, Smilgies, Detlef-M, Estevez, Jose Manuel, Bonetta, Dario, Urbanowicz, Breeanna, Ehrhardt, David, Somerville, Chris, Rose, Jocelyn, Hong, Mei, and DeBolt, Seth. Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1{sup A903V} and CESA3{sup T942I} of cellulose synthase. United States: N. p., 2012. Web. doi:10.1073/iti1312109.
Harris, Darby, Corbin, Kendall, Wang, Tuo, Gutierrez, Ryan, Bertolo, Ana, Petti, Caroalberto, Smilgies, Detlef-M, Estevez, Jose Manuel, Bonetta, Dario, Urbanowicz, Breeanna, Ehrhardt, David, Somerville, Chris, Rose, Jocelyn, Hong, Mei, & DeBolt, Seth. Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1{sup A903V} and CESA3{sup T942I} of cellulose synthase. United States. doi:10.1073/iti1312109.
Harris, Darby, Corbin, Kendall, Wang, Tuo, Gutierrez, Ryan, Bertolo, Ana, Petti, Caroalberto, Smilgies, Detlef-M, Estevez, Jose Manuel, Bonetta, Dario, Urbanowicz, Breeanna, Ehrhardt, David, Somerville, Chris, Rose, Jocelyn, Hong, Mei, and DeBolt, Seth. Sun . "Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1{sup A903V} and CESA3{sup T942I} of cellulose synthase". United States. doi:10.1073/iti1312109.
@article{osti_1044640,
title = {Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1{sup A903V} and CESA3{sup T942I} of cellulose synthase},
author = {Harris, Darby and Corbin, Kendall and Wang, Tuo and Gutierrez, Ryan and Bertolo, Ana and Petti, Caroalberto and Smilgies, Detlef-M and Estevez, Jose Manuel and Bonetta, Dario and Urbanowicz, Breeanna and Ehrhardt, David and Somerville, Chris and Rose, Jocelyn and Hong, Mei and DeBolt, Seth},
abstractNote = {The mechanisms underlying the biosynthesis of cellulose in plants are complex and still poorly understood. A central question concerns the mechanism of microfibril structure and how this is linked to the catalytic polymerization action of cellulose synthase (CESA). Furthermore, it remains unclear whether modification of cellulose microfibril structure can be achieved genetically, which could be transformative in a bio-based economy. To explore these processes in planta, we developed a chemical genetic toolbox of pharmacological inhibitors and corresponding resistance-conferring point mutations in the C-terminal transmembrane domain region of CESA1{sup A903V} and CESA3{sup T942I} in Arabidopsis thaliana. Using {sup 13}C solid-state nuclear magnetic resonance spectroscopy and X-ray diffraction, we show that the cellulose microfibrils displayed reduced width and an additional cellulose C4 peak indicative of a degree of crystallinity that is intermediate between the surface and interior glucans of wild type, suggesting a difference in glucan chain association during microfibril formation. Consistent with measurements of lower microfibril crystallinity, cellulose extracts from mutated CESA1{sup A903V} and CESA3{sup T942I} displayed greater saccharification efficiency than wild type. Using live-cell imaging to track fluorescently labeled CESA, we found that these mutants show increased CESA velocities in the plasma membrane, an indication of increased polymerization rate. Collectively, these data suggest that CESA1{sup A903V} and CESA3{sup T942I} have modified microfibril structure in terms of crystallinity and suggest that in plants, as in bacteria, crystallization biophysically limits polymerization.},
doi = {10.1073/iti1312109},
journal = {Proceedings of the National Academy of Sciences},
number = February,
volume = 2012,
place = {United States},
year = {2012},
month = {1}
}