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l-Galactose replaces l-fucose in the pectic polysaccharide rhamnogalacturonan II synthesized by the l-fucose-deficient mur1 Arabidopsis mutant

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Abstract

Arabidopsis thaliana mur1 is a dwarf mutant with altered cell-wall properties, in which l-fucose is partially replaced by l-galactose in the xyloglucan and glycoproteins. We found that the mur1 mutation also affects the primary structure of the pectic polysaccharide rhamnogalacturonan II (RG-II). In mur1 RG-II a non-reducing terminal 2-O-methyl l-galactosyl residue and a 3,4-linked l-galactosyl residue replace the non-reducing terminal 2-O-methyl l-fucosyl residue and the 3,4-linked l-fucosyl residue, respectively, that are present in wild-type RG-II. Furthermore, we found that a terminal non-reducing l-galactosyl residue, rather than the previously reported d-galactosyl residue, is present on the 2-O-methyl xylose-containing side chain of RG-II in both wild type and mur1 plants. Approximately 95% of the RG-II from wild type and mur1 plants is solubilized as a high-molecular-weight (>100 kDa) complex, by treating walls with aqueous potassium phosphate. The molecular mass of RG-II in this complex was reduced to 5–10 kDa by treatment with endopolygalacturonase, providing additional evidence that RG-II is covalently linked to homogalacturonan. The results of this study provide additional information on the structure of RG-II and the role of this pectic polysaccharide in the plant cell wall.

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Abbreviations

AIR :

Alcohol-insoluble residue

d -Gal :

d-Galactosyl

EPG :

Endopolygalacturonase

ESI–MS:

Electrospray ionization mass spectrometry

GC–MS :

Gas chromatography–mass spectrometry

1 H-NMR :

Proton nuclear magnetic resonance spectroscopy

l -Fuc :

l-Fucosyl

l -Gal :

l-Galactosyl

2-O-MeFuc :

2-O-Methyl l-fucosyl

2-O-MeGal :

2-O-Methyl l-galactosyl

2-O-MeXyl :

2-O-Methyl d-xylosyl

MWCO :

Molecular weight cut-off

RG-II :

Rhamnogalacturonan II

ppm :

Parts per million

RI :

Refractive index

SEC :

Size-exclusion chromatography

TFA :

Trifluoroacetic acid

WT :

Wild type

References

  • Baydoun EAH, Fry SC (1988) [2-3H]Mannose incorporation in cultured plant cells: Investigation of l-galactose residues of the primary cell wall. J Plant Physiol 132:484–490

    CAS  Google Scholar 

  • Bonin CP, Potter I, Vanzin GF, Reiter W-D (1997) The MUR1 gene of Arabidopsis thaliana encodes an isoform of GDP-d-mannose-4,6-dehydratase, catalyzing the first step in the de novo synthesis of GDP-l-fucose. Proc Natl Acad Sci USA 94:2085–2090

    Article  CAS  PubMed  Google Scholar 

  • Brown PH, Bellaloui N, Wimmer MA, Bassil ES, Ruiz J, Hu H, Pfeffer H, Dannel F, Romheld V (2002) Boron in plant biology. Plant Biol 4:205–223

    Article  CAS  Google Scholar 

  • Carpita NC, Gibeaut DM (1993) Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth. Plant J 3:1–30

    CAS  PubMed  Google Scholar 

  • Carpita N, McCann M (2000) The cell wall. In: Buchanan BB, Gruissem W, Jones RL (eds) Biochemistry and molecular biology of plants. American society of plant physiologists, Rockville, pp 52–109

  • Fleischer A, Titel C, Ehwald R (1998) The boron requirement and cell wall properties of growing and stationary suspension-cultured Chenopodium album L. cells. Plant Physiol 117:1401–1410

    Article  CAS  PubMed  Google Scholar 

  • Fleischer A, O’Neill MA, Ehwald R (1999) The pore size of non-graminaceous plant cell walls is rapidly decreased by borate ester cross-linking of the pectic polysaccharide rhamnogalacturonan II. Plant Physiol 121:829–838

    CAS  PubMed  Google Scholar 

  • Gerwig GJ, Kamerling JP, Vliegenthart JFG (1979) Determination of the absolute configuration of monosaccharides in complex carbohydrates by capillary G.L.C. Carbohydr Res 77:1–7

    Article  CAS  Google Scholar 

  • Hayashi T (1989) Xyloglucans in the primary cell wall. Annu Rev Plant Physiol Plant Mol Biol 40:139–168

    Google Scholar 

  • Ishii T, Matsunaga T (2001) Pectic polysaccharide rhamnogalacturonan II is covalently linked to homogalacturonan. Phytochemistry 57:969–974

    Article  CAS  PubMed  Google Scholar 

  • Ishii T, Matsunaga T, Pellerin P, O’Neill MA, Darvill A, Albersheim P (1999) The plant cell wall polysaccharide rhamnogalacturonan II self-assembles into a covalently cross-linked dimer. J Biol Chem 274:13098–13104

    CAS  PubMed  Google Scholar 

  • Iwai H, Masaoka N, Ishii T, Satoh S (2002) A pectin glucuronyltransferase gene is essential for intercellular attachment in the plant meristem. Proc Natl Acad Sci USA 99:16319–16324

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi M, Matoh T, Azuma J (1996) Two chains of rhamnogalacturonan II are cross-linked by borate-diol ester bonds in higher plant cell walls. Plant Physiol 110:1017–1020

    CAS  PubMed  Google Scholar 

  • Levy S, Staehelin LA (1992) Synthesis, assembly and function of plant cell wall macromolecules. Curr Opin Cell Biol 4:856–862

    Article  CAS  PubMed  Google Scholar 

  • Matsunaga T, Ishii T, Matsumoto S, Higuchi M, Darvill A, Albersheim P, O’Neill MA (2004) Occurrence of the primary cell wall polysaccharide rhamnogalacturonan II in pteridophytes, lycophytes, and bryophytes. Implications for the evolution of vascular plants. Plant Physiol 134:339–351

    Article  PubMed  Google Scholar 

  • McNeil M, Darvill AG, Åman P, Franzén L-E, Albersheim P (1982) Structural analysis of complex carbohydrates using high performance liquid chromatography, gas chromatography and mass spectrometry. Methods Enzymol 83:3–45

    CAS  PubMed  Google Scholar 

  • McNeil M, Darvill A, Fry SC, Albersheim P (1984) Structure and function of the primary cell walls of plants. Annu Rev Biochem 53:625–663

    CAS  PubMed  Google Scholar 

  • O’Neill MA, Eberhard S, Albersheim P, Darvill AG (2001) Requirement of borate cross-linking of cell wall rhamnogalacturonan II for Arabidopsis growth. Science 294:846–849

    Article  CAS  PubMed  Google Scholar 

  • Rayon C, Cabanes-Macheteau M, Loutelier-Bourhis C, Salliot-Maire I, Lemoine J, Reiter W-D, Lerouge P, Faye L (1999) Characterization of N-glycans from Arabidopsis. Application to a fucose-deficient mutant. Plant Physiol 119:725–733

    Article  CAS  PubMed  Google Scholar 

  • Reiter W-D (1998) Arabidopsis thaliana as a model system to study synthesis, structure, and function of the plant cell wall. Plant Physiol Biochem 36:167–176

    Article  CAS  Google Scholar 

  • Reiter W-D, Chapple CCS, Somerville CR (1993) Altered growth and cell walls in a fucose-deficient mutant of Arabidopsis. Science 261:1032–1035

    CAS  Google Scholar 

  • Reiter W-D, Chapple C, Somerville CR (1997) Mutants of Arabidopsis thaliana with altered cell wall polysaccharide composition. Plant J 12:335–345

    CAS  PubMed  Google Scholar 

  • Reuhs BL, Carlson RW, Kim JS (1993) Rhizobium fredii and Rhizobium meliloti produce 3-deoxy-d-manno-2-octulosonic acid-containing polysaccharides that are structurally analogous to group II K antigens (capsular polysaccharides) found in Escherichia coli. J Bacteriol 175:3570–3580

    CAS  PubMed  Google Scholar 

  • Reuhs BL, Kim JS, Badgett A, Carlson RW (1994) Production of cell-associated polysaccharides of Rhizobium fredii USDA205 is modulated by apigenin and host root extract. Mol Plant Microbe Interact 7:240–247

    CAS  PubMed  Google Scholar 

  • Ridley BL, O’Neill MA, Mohnen DA (2001) Pectins: structure, biosynthesis, and oligogalacturonide-related signaling. Phytochemistry 57:929–967

    CAS  PubMed  Google Scholar 

  • Roberts K (1990) Structures at the plant cell surface. Curr Opin Cell Biol 2:920–928

    CAS  PubMed  Google Scholar 

  • Spellman MW, McNeil M, Darvill AG, Albersheim P, Dell A (1983) Structure of plant cell walls. XIV. Characterization of a structurally complex heptasaccharide isolated from the pectic polysaccharide rhamnogalacturonan II. Carbohydr Res 122:131–153

    Article  CAS  Google Scholar 

  • Varner JE, Lin L-S (1989) Plant cell wall architecture. Cell 56:231–239

    CAS  PubMed  Google Scholar 

  • Weissbach A, Hurwitz J (1958) The formation of 2-keto-3-deoxyheptanoic acid in extracts of Escherichia coli B. J Biol Chem 234:705–709

    Google Scholar 

  • Whitcombe AJ, O’Neill MA, Steffan W, Darvill AG, Albersheim P (1995) Structural characterization of the pectic polysaccharide rhamnogalacturonan II. Carbohydr Res 271:15–29

    Article  CAS  PubMed  Google Scholar 

  • York WS, Darvill AG, McNeil M, Stevenson TT, Albersheim P (1985) Isolation and characterization of plant cell walls and cell wall components. Methods Enzymol 118:3–40

    Google Scholar 

  • Zablackis E, Huang J, Muller B, Darvill AG, Albersheim P (1995) Characterization of the cell-wall polysaccharides of Arabidopsis thaliana leaves. Plant Physiol 107:1129–1138

    Article  CAS  PubMed  Google Scholar 

  • Zablackis E, York WS, Pauly M, Hantus S, Reiter W-D, Chapple CCS, Albersheim P, Darvill A (1996) Substitution of l-fucose by l-galactose in cell walls of Arabidopsis mur1. Science 272:1808–1810

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by U.S. Department of Energy grants DE-FG02-96-ER20220 (to A.D.) and DE-FG05-93-ER20097 (to A.D. and P.A.) and by a National Science Foundation grant MCB-9728564 to B.L.R. We thank Prof. W.-D. Reiter of The University of Connecticut (Storrs, CO, USA) for seeds of A. thaliana ecotype Columbia and the mur1 derivative. We also thank the following from The Complex Carbohydrate Research: Dr. C. Bergmann for gifts of Aspergillus niger EPGs I and II, A. aculeatus pectin methyl esterase, and A. niger exopolygalacturonase; and Dr. A. Vernot and Prof G.-J. Boons for synthesizing 2-O-methyl l-Fuc.

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Author notes

  1. Bradley L. Reuhs

    Present address: Whistler Center for Carbohydrate Research, Food Science Department, Purdue University, West Lafayette, IN 47907-1160, USA

  2. John S. Kim

    Present address: North American Vaccine, Inc., 10150 Old Columbia Rd., Columbia, MD 21046, USA

  3. Earl Zablackis

    Present address: Aventis Pasteur, Discovery Drive, Swiftwater, PA 18370, USA

Authors and Affiliations

  1. Complex Carbohydrate Research Center and Department of Biochemistry and Molecular Biology, The University of Georgia, 315 Riverbend Road, Athens, GA 30602-4712, USA

    Bradley L. Reuhs, Joshua Glenn, Samuel B. Stephens, John S. Kim, D. Benjamin Christie, John G. Glushka, Earl Zablackis, Peter Albersheim, Alan G. Darvill & Malcolm A. O’Neill

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  1. Bradley L. Reuhs
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  2. Joshua Glenn
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  3. Samuel B. Stephens
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  10. Malcolm A. O’Neill
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Correspondence to Malcolm A. O’Neill.

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Reuhs, B.L., Glenn, J., Stephens, S.B. et al. l-Galactose replaces l-fucose in the pectic polysaccharide rhamnogalacturonan II synthesized by the l-fucose-deficient mur1 Arabidopsis mutant. Planta 219, 147–157 (2004). https://doi.org/10.1007/s00425-004-1205-x

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