The novel chloroplast outer membrane kinase KOC1 is a required component of the plastid protein import machinery

• DOI: 10.1074/jbc.m117.776468
• Publication Date: 2017-03-11T01:25:45Z
• Authors: Mónica Zufferey; Cyrille Montandon; Véronique Douet; Emilie Demarsy; Birgit Agne; Sacha Baginsky; Felix Kessler
• Keywords: Cytosol/metabolism; 0301 basic medicine; Chloroplasts; info:eu-repo/classification/ddc/580; Chloroplast Proteins/metabolism/physiology; Chromatography, Affinity; Mass Spectrometry; Chloroplast Proteins; 03 medical and health sciences; Cytosol; Protein Domains; Arabidopsis Proteins/metabolism/physiology; Escherichia coli; Chloroplasts/metabolism; Phosphorylation; Recombinant Proteins/metabolism; Chromatography; Arabidopsis Proteins; Recombinant Proteins; Protein Transport; ddc:580; Affinity; Escherichia coli/metabolism; Mutation

The biogenesis and maintenance of cell organelles such as mitochondria and chloroplasts require the import of many proteins from the cytosol, a process that is controlled by phosphorylation. In the case of chloroplasts, the import of hundreds of different proteins depends on translocons at the outer and inner chloroplast membrane (TOC and TIC, respectively) complexes. The essential protein TOC159 functions thereby as an import receptor. It has an N-terminal acidic (A-) domain that extends into the cytosol, controls receptor specificity, and is highly phosphorylated in vivo However, kinases that phosphorylate the TOC159 A-domain to enable protein import have remained elusive. Here, using co-purification with TOC159 from Arabidopsis, we discovered a novel component of the chloroplast import machinery, the regulatory kinase at the outer chloroplast membrane 1 (KOC1). We found that KOC1 is an integral membrane protein facing the cytosol and stably associates with TOC. Moreover, KOC1 phosphorylated the A-domain of TOC159 in vitro, and in mutant koc1 chloroplasts, preprotein import efficiency was diminished. koc1 Arabidopsis seedlings had reduced survival rates after transfer from the dark to the light in which protein import into plastids is required to rapidly complete chloroplast biogenesis. In summary, our data indicate that KOC1 is a functional component of the TOC machinery that phosphorylates import receptors, supports preprotein import, and contributes to efficient chloroplast biogenesis.