title: Isolation and characterization of a J domain protein that interacts with ARC1 from ornamental kale (Brassica oleracea var. acephala).
abstract: A novel J domain protein, JDP1, was isolated from ornamental kale. The C-terminus of JDP1 specifically interacted with ARC1, which has a conserved role in self-incompatibility signaling. Armadillo (ARM)-repeat containing 1 (ARC1) plays a conserved role in self-incompatibility signaling across the Brassicaceae and functions downstream of the S-locus receptor kinase. Here, we identified a J domain protein 1 (JDP1) that interacts with ARC1 using a yeast two-hybrid screen against a stigma cDNA library from ornamental kale (Brassica oleracea var. acephala). JDP1, a 38.4-kDa protein with 344 amino acids, is a member of the Hsp40 family. Fragment JDP1(57-344), originally isolated from a yeast two-hybrid cDNA library, interacted specifically with ARC1 in yeast two-hybrid assays. The N-terminus of JDP1 (JDP1(1-68)) contains a J domain, and the C-terminus of JDP1 (JDP1(69-344)) contains an X domain of unknown function. However, JDP1(69-344) was required and sufficient for interaction with ARC1 in yeast two-hybrid assays and in vitro binding assays. Moreover, JDP1(69-344) regulated the trafficking of ARC1 from the cytoplasm to the plasma membrane by interacting with ARC1 in Arabidopsis mesophyll protoplasts. Finally, Tyr(8) in the JDP1 N-terminal region was identified to be the specific site for regulating the interaction between JDP1 and BoARC1 in yeast two-hybrid assays. Possible roles of JDP1 as an interactor with ARC1 in Brassica are discussed.
authors: College of Life Sciences, Northeast Forestry University, Harbin, 150040, Heilongjiang Province, China.
authors: Xingguo
authors: X
authors: Lan
authors: Jia
authors: J
authors: Yang
authors: Mingming
authors: M
authors: Cao
authors: Yanhong
authors: Y
authors: Wang
authors: Saneyuki
authors: S
authors: Kawabata
authors: Yuhua
authors: Y
authors: Li
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D001937:Brassica / Q000235:genetics* / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D060436:Self-Incompatibility in Flowering Plants; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
chemical_list: D050956:HSP40 Heat-Shock Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; C079832:S-receptor kinase