Eukaryotic organisms are equipped with quality-control mechanisms that survey protein folding in the endoplasmic reticulum (ER) and remove non-native proteins by ER-associated degradation (ERAD). Recent research has shown that cytokinin-degrading CKX proteins are subjected to ERAD during plant development. The mechanisms of plant ERAD, including the export of substrate proteins from the ER, are not fully understood, and the molecular components involved in the ERAD of CKX are unknown. Here, we show that heavy metal-associated isoprenylated plant proteins (HIPPs) interact specifically with CKX proteins synthesized in the ER and processed by ERAD. CKX-HIPP protein complexes were detected at the ER as well as in the cytosol, suggesting that the complexes involve retrotranslocated CKX protein species. Altered CKX levels in HIPP-overexpressing and higher-order hipp mutant plants suggest that the studied HIPPs control the ERAD of CKX. Deregulation of CKX proteins caused corresponding changes in the cytokinin signaling activity and triggered typical morphological cytokinin responses. Notably, transcriptional repression of HIPP genes by cytokinin indicates a feedback regulatory mechanism of cytokinin homeostasis and signaling responses. Moreover, loss of function of HIPP genes constitutively activates the unfolded protein response and compromises the ER stress tolerance. Collectively, these results suggests that HIPPs represent novel functional components of plant ERAD.

Institute of Biology Applied Genetics Dahlem Centre of Plant Sciences Freie Universität Berlin Albrecht Thaer Weg 6 14195 Berlin Germany

Institute of Biology Applied Genetics Dahlem Centre of Plant Sciences Freie Universität Berlin Albrecht Thaer Weg 6 14195 Berlin Germany; Guangdong Provincial Key Laboratory of Conservation and Precision Utilization of Characteristic Agricultural Resources in Mountainous Areas School of Life Science of Jiaying University 514015 Mei Zhou China

Institute of Biology University of Graz Schubertstraße 51 8010 Graz Austria

Laboratory of Growth Regulators Centre of the Region Haná for Biotechnological and Agricultural Research Palacký University and Institute of Experimental Botany ASCR 78371 Olomouc Czech Republic

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