title: Small heat shock protein Hsp17.8 functions as an AKR2A cofactor in the targeting of chloroplast outer membrane proteins in Arabidopsis.
abstract: Plastid proteins that are encoded by the nuclear genome and synthesized in the cytosol undergo posttranslational targeting to plastids. Ankyrin repeat protein 2A (AKR2A) and AKR2B were recently shown to be involved in the targeting of proteins to the plastid outer envelope. However, it remains unknown whether other factors are involved in this process. In this study, we investigated a factor involved in AKR2A-mediated protein targeting to chloroplasts in Arabidopsis (Arabidopsis thaliana). Hsp17.8, a member of the class I (CI) cytosolic small heat shock proteins (sHsps), was identified in interactions with AKR2A. The interaction between Hsp17.8 and AKR2A was further confirmed by coimmunoprecipitation experiments. The carboxyl-terminal ankyrin repeat domain of AKR2A was responsible for AKR2A binding to Hsp17.8. Other CI cytosolic sHsps also interact with AKR2A to varying degrees. Additionally, Hsp17.8 binds to chloroplasts in vitro and enhances AKR2A binding to chloroplasts. HSP17.8 was expressed under normal growth conditions, and its expression increased after heat shock. Hsp17.8 exists as a dimer under normal physiological conditions, and it is converted to high oligomeric complexes, ranging from 240 kD to greater than 480 kD, after heat shock. High levels of Hsp17.8 together with AKR2A resulted in increased plastid targeting of Outer Envelope Protein7 (OEP7), a plastid outer envelope protein expressed as a green fluorescent protein fusion protein. In contrast, artificial microRNA suppression of HSP17.8 and closely related CI cytosolic sHSPs in protoplasts resulted in a reduction of OEP7:green fluorescent protein targeting to plastids. Based on these data, we propose that Hsp17.8 functions as an AKR2A cofactor in targeting membrane proteins to plastid outer membranes under normal physiological conditions.
authors: Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
authors: Dae Heon
authors: DH
authors: Kim
authors: Zheng-Yi
authors: ZY
authors: Xu
authors: Yun Jeong
authors: YJ
authors: Na
authors: Yun-Joo
authors: YJ
authors: Yoo
authors: Junho
authors: J
authors: Lee
authors: Eun-Ju
authors: EJ
authors: Sohn
authors: Inhwan
authors: I
authors: Hwang
mesh_terms: D017360:Arabidopsis / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000502:physiology*; D002736:Chloroplasts / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D050888:Heat-Shock Proteins, Small / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology*; D018832:Molecular Chaperones / Q000502:physiology*; D011485:Protein Binding
chemical_list: C451699:AKR2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D050888:Heat-Shock Proteins, Small; D008565:Membrane Proteins; D018832:Molecular Chaperones; D049452:Green Fluorescent Proteins