GUX1 and GUX2 glucuronyltransferases decorate distinct domains of glucuronoxylan with different substitution patterns

Abstract

Xylan comprises up to one-third of plant cell walls, and it influences the properties and processing of biomass. Glucuronoxylan in Arabidopsis is characterized by a linear β-(1,4)-linked backbone of xylosyl residues substituted by glucuronic acid and 4-O-methylglucuronic acid (collectively termed [Me]GlcA). The role of these substitutions remains unclear. GUX1 (glucuronic acid substitution of xylan 1) and GUX2, recently identified as glucuronyltransferases, are both required for substitution of the xylan backbone with [Me]GlcA. Here, we demonstrate clear differences in the pattern of [Me]GlcA substitution generated by each of these glucuronyltransferases. GUX1 decorates xylan with a preference for addition of [Me]GlcA at evenly spaced xylosyl residues. Intervals of eight or 10 residues dominate, but larger intervals are observed. GUX2, in contrast, produces more tightly clustered decorations with most frequent spacing of five, six or seven xylosyl residues, with no preference for odd or even spacing. Moreover, each of these GUX transferases substitutes a distinct domain of secondary cell wall xylan, which we call the major and minor domains. These major and minor xylan domains were not separable from each other by size or charge, a finding that suggests that they are tightly associated. The presence of both differently [Me]GlcA decorated domains may produce a xylan molecule that is heterogeneous in its properties. We speculate that the major and minor domains of xylan may be specialised, such as for interaction with cellulose or lignin. These findings have substantial implications for our understanding of xylan synthesis and structure, and for models of the molecular architecture of the lignocellulosic matrix of plant cell walls.