title: Galacturonosyltransferase (GAUT)1 and GAUT7 are the core of a plant cell wall pectin biosynthetic homogalacturonan:galacturonosyltransferase complex.
abstract: Plant cell wall pectic polysaccharides are arguably the most complex carbohydrates in nature. Progress in understanding pectin synthesis has been slow due to its complex structure and difficulties in purifying and expressing the low-abundance, Golgi membrane-bound pectin biosynthetic enzymes. Arabidopsis galacturonosyltransferase (GAUT) 1 is an α-1,4-galacturonosyltransferase (GalAT) that synthesizes homogalacturonan (HG), the most abundant pectic polysaccharide. We now show that GAUT1 functions in a protein complex with the homologous GAUT7. Surprisingly, although both GAUT1 and GAUT7 are type II membrane proteins with single N-terminal transmembrane-spanning domains, the N-terminal region of GAUT1, including the transmembrane domain, is cleaved in vivo. This raises the question of how the processed GAUT1 is retained in the Golgi, the site of HG biosynthesis. We show that the anchoring of GAUT1 in the Golgi requires association with GAUT7 to form the GAUT1:GAUT7 complex. Proteomics analyses also identified 12 additional proteins that immunoprecipitate with the GAUT1:GAUT7 complex. This study provides conclusive evidence that the GAUT1:GAUT7 complex is the catalytic core of an HG:GalAT complex and that cell wall matrix polysaccharide biosynthesis occurs via protein complexes. The processing of GAUT1 to remove its N-terminal transmembrane domain and its anchoring in the Golgi by association with GAUT7 provides an example of how specific catalytic domains of plant cell wall biosynthetic glycosyltransferases could be assembled into protein complexes to enable the synthesis of the complex and developmentally and environmentally plastic plant cell wall.
authors.affiliation: Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USA.
authors.forename: Melani A
authors.initials: MA
authors.lastname: Atmodjo
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Sakuragi
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhu
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Burrell
authors.forename: Sushree S
authors.initials: SS
authors.lastname: Mohanty
authors.forename: James A
authors.initials: JA
authors.lastname: Atwood
authors.forename: Ron
authors.initials: R
authors.lastname: Orlando
authors.forename: Henrik V
authors.initials: HV
authors.lastname: Scheller
authors.forename: Debra
authors.initials: D
authors.lastname: Mohnen
mesh_terms: D017360:Arabidopsis / Q000166:cytology* / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002473:Cell Wall / Q000201:enzymology*; D014453:Glucuronosyltransferase; D006056:Golgi Apparatus / Q000201:enzymology; D047468:Immunoprecipitation; D008954:Models, Biological; D046912:Multiprotein Complexes / Q000378:metabolism; D010368:Pectins / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D040901:Proteomics; D013379:Substrate Specificity
chemical_list: D029681:Arabidopsis Proteins; D046912:Multiprotein Complexes; D010368:Pectins; D014453:Glucuronosyltransferase; C569601:galacturonosyltransferase 1, Arabidopsis; C569602:galacturonosyltransferase 7, Arabidopsis; C003181:polygalacturonic acid