skip to main content

DOE PAGESDOE PAGES

Title: Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity

Here, cellulose microfibrils are synthesized by membrane-embedded cellulose synthesis complexes (CSCs), currently modeled as hexamers of cellulose synthase (CESA) trimers. The three paralogous CESAs involved in secondary cell wall (SCW) cellulose biosynthesis in Arabidopsis (CESA4, CESA7, CESA8) are similar, but nonredundant, with all three isoforms required for assembly and function of the CSC. The molecular basis of protein–protein recognition among the isoforms is not well understood. To investigate the locations of the interfaces that are responsible for isoform recognition, we swapped three domains between the Arabidopsis CESAs required for SCW synthesis (CESA4, CESA7, and CESA8): N-terminus, central domain containing the catalytic core, and C-terminus. Chimeric genes with all pairwise permutations of the domains were tested for in vivo functionality within knockout mutant backgrounds of cesa4, cesa7, and cesa8. Immunoblotting with isoform-specific antibodies confirmed the anticipated protein expression in transgenic plants. The percent recovery of stem height and crystalline cellulose content was assayed, as compared to wild type, the mutant background lines, and other controls. Retention of the native central domain was sufficient for CESA8 chimeras to function, with neither its N-terminal nor C-terminal domains required. The C-terminal domain is required for class-specific function of CESA4 and CESA7, and CESA7 alsomore » requires its own N-terminus. Across all isoforms, the results indicate that the central domain, as well as the N- and C-terminal regions, contributes to class-specific function variously in Arabidopsis CESA4, CESA7, and CESA8.« less
Authors:
 [1] ;  [1] ;  [2] ;  [3] ;  [1]
  1. Pennsylvania State Univ., University Park, PA (United States)
  2. Univ. of Rhode Island, Kingston, RI (United States)
  3. North Carolina State Univ., Raleigh, NC (United States)
Publication Date:
Grant/Contract Number:
SC0001090
Type:
Published Article
Journal Name:
Plant Direct
Additional Journal Information:
Journal Volume: 2; Journal Issue: 7; Related Information: CLSF partners with Pennsylvania State University (lead); North Carolina State University; University of Rhode Island; Virginia Tech University; Journal ID: ISSN 2475-4455
Publisher:
Wiley and American Society of Plant Biologists and Society for Experimental Biology
Research Org:
Energy Frontier Research Centers (EFRC), Washington, D.C. (United States). Center for Lignocellulose Structure and Formation (CLSF)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; biofuels (including algae and biomass); bio-inspired; membrane; carbon sequestration; materials and chemistry by design; synthesis (self-assembly)
OSTI Identifier:
1459680
Alternate Identifier(s):
OSTI ID: 1459681; OSTI ID: 1470661

Hill, Jr., Joseph Lee, Hill, Ashley Nicole, Roberts, Alison W., Haigler, Candace H., and Tien, Ming. Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity. United States: N. p., Web. doi:10.1002/pld3.61.
Hill, Jr., Joseph Lee, Hill, Ashley Nicole, Roberts, Alison W., Haigler, Candace H., & Tien, Ming. Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity. United States. doi:10.1002/pld3.61.
Hill, Jr., Joseph Lee, Hill, Ashley Nicole, Roberts, Alison W., Haigler, Candace H., and Tien, Ming. 2018. "Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity". United States. doi:10.1002/pld3.61.
@article{osti_1459680,
title = {Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity},
author = {Hill, Jr., Joseph Lee and Hill, Ashley Nicole and Roberts, Alison W. and Haigler, Candace H. and Tien, Ming},
abstractNote = {Here, cellulose microfibrils are synthesized by membrane-embedded cellulose synthesis complexes (CSCs), currently modeled as hexamers of cellulose synthase (CESA) trimers. The three paralogous CESAs involved in secondary cell wall (SCW) cellulose biosynthesis in Arabidopsis (CESA4, CESA7, CESA8) are similar, but nonredundant, with all three isoforms required for assembly and function of the CSC. The molecular basis of protein–protein recognition among the isoforms is not well understood. To investigate the locations of the interfaces that are responsible for isoform recognition, we swapped three domains between the Arabidopsis CESAs required for SCW synthesis (CESA4, CESA7, and CESA8): N-terminus, central domain containing the catalytic core, and C-terminus. Chimeric genes with all pairwise permutations of the domains were tested for in vivo functionality within knockout mutant backgrounds of cesa4, cesa7, and cesa8. Immunoblotting with isoform-specific antibodies confirmed the anticipated protein expression in transgenic plants. The percent recovery of stem height and crystalline cellulose content was assayed, as compared to wild type, the mutant background lines, and other controls. Retention of the native central domain was sufficient for CESA8 chimeras to function, with neither its N-terminal nor C-terminal domains required. The C-terminal domain is required for class-specific function of CESA4 and CESA7, and CESA7 also requires its own N-terminus. Across all isoforms, the results indicate that the central domain, as well as the N- and C-terminal regions, contributes to class-specific function variously in Arabidopsis CESA4, CESA7, and CESA8.},
doi = {10.1002/pld3.61},
journal = {Plant Direct},
number = 7,
volume = 2,
place = {United States},
year = {2018},
month = {7}
}

Works referenced in this record:

Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana
journal, August 2003
  • Alonso, J. M.; Stepanova, Anna N.; Leisse, Thomas J.
  • Science, Vol. 301, Issue 5633, p. 653-657
  • DOI: 10.1126/science.1086391

Collapsed Xylem Phenotype of Arabidopsis Identifies Mutants Deficient in Cellulose Deposition in the Secondary Cell Wall
journal, May 1997
  • Turner, S. R.; Somerville, C. R.
  • The Plant Cell, Vol. 9, Issue 5, p. 689-701
  • DOI: 10.1105/tpc.9.5.689

Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana
journal, December 1998

Multiple sequence alignment with hierarchical clustering
journal, November 1988