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Title: The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2

Abstract

Traditional marker-based mapping and next-generation sequencing was used to determine that the Arabidopsis (Arabidopsis thaliana) low cell wall arabinose mutant murus5 (mur5) encodes a defective allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2 (RGP2). Marker analysis of 13 F2 confirmed mutant progeny from a recombinant mapping population gave a rough map position on the upper arm of chromosome 5, and deep sequencing of DNA from these 13 lines gave five candidate genes with G→A (C→T) transitions predicted to result in amino acid changes. Of these five, only insertional mutant alleles of RGP2, a gene that encodes a UDP-arabinose mutase that interconverts UDP-arabinopyranose and UDP-arabinofuranose, exhibited the low cell wall arabinose phenotype. The identities of mur5 and two SALK insertional alleles were confirmed by allelism tests and overexpression of wild-type RGP2 complementary DNA placed under the control of the 35S promoter in the three alleles. The mur5 mutation results in the conversion of cysteine-257 to tyrosine-257 within a conserved hydrophobic cluster predicted to be distal to the active site and essential for protein stability and possible heterodimerization with other isoforms of RGP.

Authors:
 [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3]; ORCiD logo [4]; ORCiD logo [1];  [1]; ORCiD logo [1];  [1]; ORCiD logo [4]; ORCiD logo [1]; ORCiD logo [1]
  1. Purdue Univ., West Lafayette, IN (United States)
  2. Univ. de Picardie Jules Verne, Amiens (France)
  3. Heidelberg Inst. für Theoretische Studien, Molecular Biomechanics, Heidelberg (Germany)
  4. Univ. of Connecticut, Storrs, CT (United States). Dept. of Molecular and Cell Biology
Publication Date:
Research Org.:
Energy Frontier Research Centers (EFRC) (United States). Center for Direct Catalytic Conversion of Biomass to Biofuels (C3Bio)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1388829
Grant/Contract Number:  
SC000997
Resource Type:
Accepted Manuscript
Journal Name:
Plant Physiology (Bethesda)
Additional Journal Information:
Journal Name: Plant Physiology (Bethesda); Journal Volume: 171; Journal Issue: 3; Related Information: C3Bio partners with Purdue University (lead); Argonne National Laboratory; National Renewable Energy Laboratory; Northeastern University; University of Tennessee; Journal ID: ISSN 0032-0889
Publisher:
American Society of Plant Biologists
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; catalysis (homogeneous); catalysis (heterogeneous); biofuels (including algae and biomass); bio-inspired; materials and chemistry by design; synthesis (self-assembly); synthesis (scalable processing)

Citation Formats

Dugard, Christopher K., Mertz, Rachel A., Rayon, Catherine, Mercadante, Davide, Hart, Christopher, Benatti, Matheus R., Olek, Anna T., SanMiguel, Phillip J., Cooper, Bruce R., Reiter, Wolf-Dieter, McCann, Maureen C., and Carpita, Nicholas C. The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2. United States: N. p., 2016. Web. doi:10.1104/pp.15.01922.
Dugard, Christopher K., Mertz, Rachel A., Rayon, Catherine, Mercadante, Davide, Hart, Christopher, Benatti, Matheus R., Olek, Anna T., SanMiguel, Phillip J., Cooper, Bruce R., Reiter, Wolf-Dieter, McCann, Maureen C., & Carpita, Nicholas C. The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2. United States. doi:10.1104/pp.15.01922.
Dugard, Christopher K., Mertz, Rachel A., Rayon, Catherine, Mercadante, Davide, Hart, Christopher, Benatti, Matheus R., Olek, Anna T., SanMiguel, Phillip J., Cooper, Bruce R., Reiter, Wolf-Dieter, McCann, Maureen C., and Carpita, Nicholas C. Sat . "The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2". United States. doi:10.1104/pp.15.01922. https://www.osti.gov/servlets/purl/1388829.
@article{osti_1388829,
title = {The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2},
author = {Dugard, Christopher K. and Mertz, Rachel A. and Rayon, Catherine and Mercadante, Davide and Hart, Christopher and Benatti, Matheus R. and Olek, Anna T. and SanMiguel, Phillip J. and Cooper, Bruce R. and Reiter, Wolf-Dieter and McCann, Maureen C. and Carpita, Nicholas C.},
abstractNote = {Traditional marker-based mapping and next-generation sequencing was used to determine that the Arabidopsis (Arabidopsis thaliana) low cell wall arabinose mutant murus5 (mur5) encodes a defective allele of REVERSIBLY GLYCOSYLATED POLYPEPTIDE2 (RGP2). Marker analysis of 13 F2 confirmed mutant progeny from a recombinant mapping population gave a rough map position on the upper arm of chromosome 5, and deep sequencing of DNA from these 13 lines gave five candidate genes with G→A (C→T) transitions predicted to result in amino acid changes. Of these five, only insertional mutant alleles of RGP2, a gene that encodes a UDP-arabinose mutase that interconverts UDP-arabinopyranose and UDP-arabinofuranose, exhibited the low cell wall arabinose phenotype. The identities of mur5 and two SALK insertional alleles were confirmed by allelism tests and overexpression of wild-type RGP2 complementary DNA placed under the control of the 35S promoter in the three alleles. The mur5 mutation results in the conversion of cysteine-257 to tyrosine-257 within a conserved hydrophobic cluster predicted to be distal to the active site and essential for protein stability and possible heterodimerization with other isoforms of RGP.},
doi = {10.1104/pp.15.01922},
journal = {Plant Physiology (Bethesda)},
number = 3,
volume = 171,
place = {United States},
year = {2016},
month = {7}
}

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Figures / Tables:

Figure 1 Figure 1: Characterization of the low cell wall Ara phenotype. A, Comparison of changes in Ara mol % of total trifluoroacetic acid (TFA)-soluble cell wall monosaccharide fractions from rosette leaves of Columbia-0 (Col-0), mur4, mur5, and mur6 at 14, 25, and 35 d of growth. B, Comparison of mol %more » of TFA-soluble cell wall monosaccharide fractions (excluding Glc) from Col-0 and mur5 at 35 d of growth. C, Relative cell wall content of Ara in Col-0 and mutant plants fed exogenous Ara for 28 d.« less

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