Essential Amino Acids in the Plant-Conserved and Class-Specific Regions of Cellulose Synthases: Article No. kiac479

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

Research output: Contribution to journalArticlepeer-review

2 Scopus Citations

Abstract

The Plant-Conserved Region (P-CR) and the Class-Specific Region (CSR) are two plant-unique sequences in the catalytic core of cellulose synthases (CESAs) for which specific functions have not been established. Here, we used site-directed mutagenesis to replace amino acids and motifs within these sequences predicted to be essential for assembly and function of CESAs. We developed an in vivo method to determine the ability of mutated CesA1 transgenes to complement an Arabidopsis (Arabidopsis thaliana) temperature-sensitive root-swelling1 (rsw1) mutant. Replacement of a Cys residue in the CSR, which blocks dimerization in vitro, rendered the AtCesA1 transgene unable to complement the rsw1 mutation. Examination of the CSR sequences from 33 diverse angiosperm species showed domains of high-sequence conservation in a class-specific manner but with variation in the degrees of disorder, indicating a nonredundant role of the CSR structures in different CESA isoform classes. The Cys residue essential for dimerization was not always located in domains of intrinsic disorder. Expression of AtCesA1 transgene constructs, in which Pro417 and Arg453 were substituted for Ala or Lys in the coiled-coil of the P-CR, were also unable to complement the rsw1 mutation. Despite an expected role for Arg457 in trimerization of CESA proteins, AtCesA1 transgenes with Arg457Ala mutations were able to fully restore the wild-type phenotype in rsw1. Our data support that Cys662 within the CSR and Pro417 and Arg453 within the P-CR of Arabidopsis CESA1 are essential residues for functional synthase complex formation, but our data do not support a specific role for Arg457 in trimerization in native CESA complexes.
Original languageAmerican English
Pages (from-to)142-160
Number of pages19
JournalPlant Physiology
Volume191
Issue number1
DOIs
StatePublished - 2023

NREL Publication Number

  • NREL/JA-2700-83202

Keywords

  • cellulose biosynthesis
  • cellulose synthase structure
  • intrinsic disorder
  • mutation analysis
  • protein structure modeling
  • proteomics
  • recombinant DNA

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