title: Identification of the region of rice 13 kDa prolamin essential for the formation of ER-derived protein bodies using a heterologous expression system.
abstract: Cereal prolamins, which are alcohol-soluble seed storage proteins, can induce ER-derived protein bodies (PBs) in heterologous tissue. Like maize and wheat prolamins, rice prolamins can form ER-derived PBs, but the region of mature polypeptides that is essential for PB formation has not been identified. In this study, we examined the formation mechanisms of ER-derived PB-like structures by expressing rice 13 kDa prolamin-deletion mutants fused to green fluorescent protein (GFP) in heterologous tissues such as yeast. The 13 kDa prolamin-GFP fusion protein was stably accumulated in transgenic yeast and formed an ER-derived PB-like structure. In contrast, rice α-globulin-GFP fusion protein was transported to vacuoles. In addition, the middle and COOH-terminal regions of 13 kDa prolamin formed ER-derived PB-like structures, whereas the NH2-terminal region of 13 kDa prolamin did not form such structures. These results suggest that the middle and COOH-terminal regions of 13 kDa prolamin can be retained and thus can induce ER-derived PB in yeast.
authors.affiliation: a Laboratory of Genetic Engineering, Graduate School of Life and Environmental Sciences , Kyoto Prefectural University , Kyoto , Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Masumura
authors.forename: Takanari
authors.initials: T
authors.lastname: Shigemitsu
authors.forename: Shigeto
authors.initials: S
authors.lastname: Morita
authors.forename: Shigeru
authors.initials: S
authors.lastname: Satoh
mesh_terms: D000510:Alpha-Globulins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D055315:Prolamins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics* / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
chemical_list: D000510:Alpha-Globulins; D055315:Prolamins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: ER-derived PB; GFP; prolamin; rice