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

Joseph Lee Hill, Ashley Nicole Hill, Alison W. Roberts, Candace H. Haigler, Ming Tien

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article numbere00061
JournalPlant Direct
Volume2
Issue number7
DOIs
StatePublished - Jul 1 2018

Fingerprint

cellulose synthase
Arabidopsis
Cellulose
cellulose
Protein Isoforms
Cell Wall
synthesis
cell walls
Microfibrils
knockout mutants
chimerism
Genetically Modified Plants
Biosynthesis
chimera
transgenic plant
immunoblotting
Immunoblotting
transgenic plants
Catalytic Domain
protein synthesis

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Ecology
  • Ecology, Evolution, Behavior and Systematics
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Hill, 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. In: Plant Direct. 2018 ; Vol. 2, No. 7.
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Domain swaps of Arabidopsis secondary wall cellulose synthases to elucidate their class specificity. / Hill, Joseph Lee; Hill, Ashley Nicole; Roberts, Alison W.; Haigler, Candace H.; Tien, Ming.

In: Plant Direct, Vol. 2, No. 7, e00061, 01.07.2018.

Research output: Contribution to journalArticle

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