The Cell Wall Arabinose-Deficient Arabidopsis thaliana Mutant murus5 Encodes a Defective Allele of REVERSIBLY
Christopher K Dugard1, Rachel A Mertz1, Catherine Rayon1
1Department of Botany and Plant Pathology (C.K.D., R.A.M., A.T.O., N.C.C.), Department of Biological Sciences (M.R.B., M.C.M., N.C.C.), Bindley Bioscience Center (B.R.C., M.C.M., N.C.C.), and Department of Horticulture and Landscape Architecture (P.J.S.), Purdue University, West Lafayette, Indiana 47907-2054;Université de Picardie Jules Verne, EA 3900-BIOPI, 80039 Amiens, France (C.R.);Heidelberg Institut für Theoretische Studien, Molecular Biomechanics, 69118 Heidelberg, Germany (D.M.); andDepartment of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut 06269 (C.H., W.-D.R.).
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.
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