MassIVE MSV000089874

Partial Public

Amino acids essential for function in the Plant-Conserved and Class-Specific Regions of plant cellulose synthase

Subscribe Comment Add Reanalysis

Description

The Plant-Conserved Region (P-CR) and the Class-Specific Region (CSR) are two unique sequences inserted into the catalytic core of plant cellulose synthases. Either or both of these domains might promote assembly of the synthase complex. From structural studies of these domains, we have identified candidate amino acids and motifs that might be essential for assembly of the complex or catalytic function. We used site-directed mutagenesis to replace them in the CSR and P-CR in an Arabidopsis AtCesA1 and tested the ability of the mutated CesA1 transgenes to complement an Arabidopsis temperature-sensitive root-swelling1 (rsw1) mutant and restore the wild-type phenotype. Elimination of conserved basic and acidic motifs characteristic of the CSR in AtCesA1 constructs were able to complement wild-type growth and cellulose synthesis in rsw1. However, transgenes that replaced Cys662 with Ala failed to complement, indicating an essential function for this residue. Replacement of the cognate Cys662 with Ala in a recombinant rice CESA8 catalytic domain eliminated the ability of the recombinant protein to dimerize. Expression of transgene constructs in which of Pro417 and Arg453 were substituted for Ala or Lys in the coiled-coil of the P-CR also were unable to complement the rsw1 mutation. However, mutation of Arg457, residue predicted to function in trimerization of reconstituted poplar CESA8, fully restored the wild-type phenotype in transgenic rsw1. Taken together, these results indicate that Cys residues are essential for CesA function, most likely through their role in dimerization of CSR domains. Although evidence supports roles for both the CSR and P-CR in oligomerization of CesAs, our data do not support a role for Arg457 in trimerization in the native CesA complex. Examination of the 3-dimensional structure of a poplar CESA8 revealed other amino acids that might stabilize trimer structures in the absence of Arg457. [doi:10.25345/C5VH5CN88] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Cellulose synthase, Arabidopsis, proteomics, temperature sensitive mutants

Contact

Principal Investigators:
(in alphabetical order) 		Nicholas Carpita, Biosciences Center, National Renewable Energy Laboratory, United States
Submitting User: uma_aryal
Number of Files:
Total Size:
Spectra:
Subscribers:
 
Owner Reanalyses
Experimental Design
    Conditions:
    Biological Replicates:
    Technical Replicates:
 
Identification Results
    Proteins (Human, Remapped):
    Proteins (Reported):
    Peptides:
    Variant Peptides:
    PSMs:
 
Quantification Results
    Differential Proteins:
    Quantified Proteins:
 
Browse Dataset Files
 
FTP Download Link (click to copy):

Species

Instrument

Modifications

- Dataset Reanalyses

+ Dataset History

Click here to queue conversion of this dataset's submitted spectrum files to open formats (e.g. mzML). This process may take some time.

When complete, the converted files will be available in the "ccms_peak" subdirectory of the dataset's FTP space (accessible via the "FTP Download" link to the right).
Number of distinct conditions across all analyses (original submission and reanalyses) associated with this dataset.

Distinct condition labels are counted across all files submitted in the "Metadata" category having a "Condition" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct biological replicates across all analyses (original submission and reanalyses) associated with this dataset.

Distinct replicate labels are counted across all files submitted in the "Metadata" category having a "BioReplicate" or "Replicate" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct technical replicates across all analyses (original submission and reanalyses) associated with this dataset.

The technical replicate count is defined as the maximum number of times any one distinct combination of condition and biological replicate was analyzed across all files submitted in the "Metadata" category. In the case of fractionated experiments, only the first fraction is considered.

"N/A" means no results of this type were submitted.
Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

"N/A" means no results of this type were submitted.
Number of distinct protein accessions reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct unmodified peptide sequences reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct peptide sequences (including modified variants or peptidoforms) reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Total number of peptide-spectrum matches (i.e. spectrum identifications) reported across all analyses (original submission and reanalyses) associated with this dataset.

"N/A" means no results of this type were submitted.
Number of distinct proteins quantified across all analyses (original submission and reanalyses) associated with this dataset.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
Number of distinct proteins found to be differentially abundant in at least one comparison across all analyses (original submission and reanalyses) associated with this dataset.

A protein is differentially abundant if its change in abundance across conditions is found to be statistically significant with an adjusted p-value <= 0.05 and lists no issues associated with statistical tests for differential abundance.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
This dataset may not contain all raw spectra data as originally deposited in PRIDE. It has been imported to MassIVE for reanalysis purposes, so its spectra data here may consist solely of processed peak lists suitable for reanalysis with most software.