title: Identification of an additional protein involved in mannan biosynthesis.
abstract: Galactomannans comprise a β-1,4-mannan backbone substituted with α-1,6-galactosyl residues. Genes encoding the enzymes that are primarily responsible for backbone synthesis and side-chain addition of galactomannans were previously identified and characterized. To identify additional genes involved in galactomannan biosynthesis, we previously performed deep EST profiling of fenugreek (Trigonella foenum-graecum L.) seed endosperm, which accumulates large quantities of galactomannans as a reserve carbohydrate during seed development. One of the candidate genes encodes a protein that is likely to be a glycosyltransferase. Because this protein is involved in mannan biosynthesis, we named it 'mannan synthesis-related' (MSR). Here, we report the characterization of a fenugreek MSR gene (TfMSR) and its two Arabidopsis homologs, AtMSR1 and AtMSR2. TfMSR was highly and specifically expressed in the endosperm. TfMSR, AtMSR1 and AtMSR2 proteins were all determined to be localized to the Golgi by fluorescence confocal microscopy. The level of mannosyl residues in stem glucomannans decreased by approximately 40% for Arabidopsis msr1 single T-DNA insertion mutants and by more than 50% for msr1 msr2 double mutants, but remained unchanged for msr2 single mutants. In addition, in vitro mannan synthase activity from the stems of msr1 single and msr1 msr2 double mutants also decreased. Expression of AtMSR1 or AtMSR2 in the msr1 msr2 double mutant completely or partially restored mannosyl levels. From these results, we conclude that the MSR protein is important for mannan biosynthesis, and offer some ideas about its role.
authors.affiliation: Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, MI, 48824, USA.|Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, MI, 48824, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QW, UK.
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Mortimer
authors.affiliation: Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, MI, 48824, USA.|Department of Plant Biology, Michigan State University, East Lansing, MI, 48824, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Davis
authors.affiliation: Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QW, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Dupree
authors.affiliation: Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, MI, 48824, USA.|Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, MI, 48824, USA.|Department of Plant Biology, Michigan State University, East Lansing, MI, 48824, USA.|Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Keegstra
mesh_terms: D056625:Endosperm / Q000378:metabolism; D017343:Genes, Plant / Q000502:physiology; D006056:Golgi Apparatus / Q000378:metabolism; D008351:Mannans / Q000096:biosynthesis*; D008364:Mannosyltransferases / Q000378:metabolism / Q000502:physiology; D008861:Microsomes / Q000378:metabolism; D010940:Plant Proteins / Q000502:physiology; D029910:Trigonella / Q000378:metabolism*
chemical_list: D008351:Mannans; D010940:Plant Proteins; D008364:Mannosyltransferases; C021542:mannan synthetase