Asparagus IRX9, IRX10, and IRX14A Are Components of an Active Xylan Backbone Synthase Complex that Forms in the Golgi Apparatus  . Issue 1 (7th March 2016)

• Record Type: Journal Article
• Authors: Zeng, Wei; Lampugnani, Edwin R.; Picard, Kelsey L.; Song, Lili; Wu, Ai-Min; Farion, Isabela M.; Zhao, Jia; Ford, Kris; Doblin, Monika S.; Bacic, Antony
• Is Part Of: Plant physiology. Volume 171:Issue 1(2016)
• Page Start: 93
• Page End: 109
• Publication Date: 2016-03-07
• Subjects: Plant physiology -- Periodicals; Botany -- Periodicals; Periodicals; Electronic journals; 571.2
• DOI: 10.1104/pp.15.01919
• Languages: English
• ISSNs: 0032-0889

Abstract

Abstract : Xylan biosynthesis in the Golgi apparatus is affected by IRX9, IRX10, and IRX14A in Asparagus. Abstract: Heteroxylans are abundant components of plant cell walls and provide important raw materials for the food, pharmaceutical, and biofuel industries. A number of studies in Arabidopsis ( Arabidopsis thaliana ) have suggested that the IRREGULAR XYLEM9 (IRX9), IRX10, and IRX14 proteins, as well as their homologs, are involved in xylan synthesis via a Golgi-localized complex termed the xylan synthase complex (XSC ). However, both the biochemical and cell biological research lags the genetic and molecular evidence. In this study, we characterized garden asparagus ( Asparagus officinalis ) stem xylan biosynthesis genes ( AoIRX9, AoIRX9L, AoIRX10, AoIRX14A, and AoIRX14B ) by heterologous expression in Nicotiana benthamiana . We reconstituted and partially purified an active XSC and showed that three proteins, AoIRX9, AoIRX10, and AoIRX14A, are necessary for xylan xylosyltranferase activity in planta. To better understand the XSC structure and its composition, we carried out coimmunoprecipitation and bimolecular fluorescence complementation analysis to show the molecular interactions between these three IRX proteins. Using a site-directed mutagenesis approach, we showed that the DxD motifs of AoIRX10 and AoIRX14A are crucial for the catalytic activity. These data provide, to our knowledge, the first lines of biochemical and cell biological evidence that AoIRX9, AoIRX10, and AoIRX14A are core components of a Golgi-localized XSC, each with distinct roles for effective heteroxylan biosynthesis.