Amino Acids Essential for the Assembly of Cellulose Synthase Complexes

Anna Olek, Phillip Rushton, Daisuke Kihara, Uma Aryal, Peter Ciesielski, Bryon Donohoe, Zicong Zhang, Cynthia Stauffacher, Maureen McCann, Nicholas Carpita

Research output: NRELPresentation


Over 40 years ago, Andrew Staehelin's group showed that plant cellulose microfibrils are synthesized by protein complexes arranged in hexagonal arrays called 'particle rosettes'. Plant cellulose synthases (CESAs) differ from their bacterial ancestral forms by insertion of three unique sequences responsible for the assembly into compact multimeric units that aggregate further into rosettes. We used site-directed mutagenesis to replace amino acids within these sequences predicted to be essential for assembly and developed an in vivo method to determine the ability of mutated CesA1 transgenes to complement an Arabidopsis temperature-sensitive root-swelling1 (rsw1) mutant. Replacement of a Cys residue in the Class-Specific Region (CSR) or Pro417 and Arg453 of the Plant-Conserved Region (P-CR) rendered an AtCesA1 transgene unable to complement the rsw1 mutation. Despite an expected role for Arg457 in the trimerization of CESA proteins, AtCesA1 transgenes with Arg457Ala mutations were able to fully restore the wild-type phenotype in rsw1. Staehelin observed a third order of assembly in Micrasterias dendiculata, where hundreds of rosettes form large hexagonal arrays during the synthesis of secondary wall cellulose macrofibrils. We found by SEM of cytosolic faces of Micrasterias cell fragments that these arrays are associated with fields of regularly spaced slime secretion pore complexes.
Original languageAmerican English
Number of pages13
StatePublished - 2023

Publication series

NamePresented at the XVI Plant Cell Wall Meeting, 18-22 June 2023, Malaga, Spain

NREL Publication Number

  • NREL/PR-2700-86384


  • arabidopsis
  • cellulose synthase complexes
  • cellulose synthesis
  • class-specific region
  • genetic complementation
  • micrasterias
  • plant-conserved region
  • secretion bodies


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