title: Brittle Culm1, a COBRA-like protein, functions in cellulose assembly through binding cellulose microfibrils.
abstract: Cellulose represents the most abundant biopolymer in nature and has great economic importance. Cellulose chains pack laterally into crystalline forms, stacking into a complicated crystallographic structure. However, the mechanism of cellulose crystallization is poorly understood. Here, via functional characterization, we report that Brittle Culm1 (BC1), a COBRA-like protein in rice, modifies cellulose crystallinity. BC1 was demonstrated to be a glycosylphosphatidylinositol (GPI) anchored protein and can be released into cell walls by removal of the GPI anchor. BC1 possesses a carbohydrate-binding module (CBM) at its N-terminus. In vitro binding assays showed that this CBM interacts specifically with crystalline cellulose, and several aromatic residues in this domain are essential for binding. It was further demonstrated that cell wall-localized BC1 via the CBM and GPI anchor is one functional form of BC1. X-ray diffraction (XRD) assays revealed that mutations in BC1 and knockdown of BC1 expression decrease the crystallite width of cellulose; overexpression of BC1 and the CBM-mutated BC1s caused varied crystallinity with results that were consistent with the in vitro binding assay. Moreover, interaction between the CBM and cellulose microfibrils was largely repressed when the cell wall residues were pre-stained with two cellulose dyes. Treating wild-type and bc1 seedlings with the dyes resulted in insensitive root growth responses in bc1 plants. Combined with the evidence that BC1 and three secondary wall cellulose synthases (CESAs) function in different steps of cellulose production as revealed by genetic analysis, we conclude that BC1 modulates cellulose assembly by interacting with cellulose and affecting microfibril crystallinity.
authors.affiliation: State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Lifeng
authors.initials: L
authors.lastname: Liu
authors.forename: Keke
authors.initials: K
authors.lastname: Shang-Guan
authors.forename: Baocai
authors.initials: B
authors.lastname: Zhang
authors.forename: Xiangling
authors.initials: X
authors.lastname: Liu
authors.forename: Meixian
authors.initials: M
authors.lastname: Yan
authors.forename: Lanjun
authors.initials: L
authors.lastname: Zhang
authors.forename: Yanyun
authors.initials: Y
authors.lastname: Shi
authors.forename: Mu
authors.initials: M
authors.lastname: Zhang
authors.forename: Qian
authors.initials: Q
authors.lastname: Qian
authors.forename: Jiayang
authors.initials: J
authors.lastname: Li
authors.forename: Yihua
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D002473:Cell Wall / Q000235:genetics / Q000378:metabolism; D002482:Cellulose / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D020894:Microfibrils / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D012275:Oryza / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D017261:Glycosylphosphatidylinositols; D002482:Cellulose; D005964:Glucosyltransferases; C478648:cellulose synthase