The COP9 Signalosome regulates seed germination by facilitating protein degradation of RGL2 and ABI5

The control of seed germination and seed dormancy are critical for the successful propagation of plant species, and are important agricultural traits. Seed germination is tightly controlled by the balance of gibberellin (GA) and abscisic acid (ABA), and is influenced by environmental factors. The COP9 Signalosome (CSN) is a conserved multi-subunit protein complex that is best known as a regulator of the Cullin-RING family of ubiquitin E3 ligases (CRLs). Multiple viable mutants of the CSN showed poor germination, except for csn5b-1. Detailed analyses showed that csn1-10 has a stronger seed dormancy, while csn5a-1 mutants exhibit retarded seed germination in addition to hyperdormancy. Both csn5a-1 and csn1-10 plants show defects in the timely removal of the germination inhibitors: RGL2, a repressor of GA signaling, and ABI5, an effector of ABA responses. We provide genetic evidence to demonstrate that the germination phenotype of csn1-10 is caused by over-accumulation of RGL2, a substrate of the SCF (CRL1) ubiquitin E3 ligase, while the csn5a-1 phenotype is caused by over-accumulation of RGL2 as well as ABI5. The genetic data are consistent with the hypothesis that CSN5A regulates ABI5 by a mechanism that may not involve CSN1. Transcriptome analyses suggest that CSN1 has a more prominent role than CSN5A during seed maturation, but CSN5A plays a more important role than CSN1 during seed germination, further supporting the functional distinction of these two CSN genes. Our study delineates the molecular targets of the CSN complex in seed germination, and reveals that CSN5 has additional functions in regulating ABI5, thus the ABA signaling pathway.
Author summary The control of seed germination and seed dormancy are critical for successful propagation of plant species, and manipulation of these processes is important for agriculture. The COP9 Signalosome (CSN) is a multi-subunit protein complex that regulates proteasome-mediated protein degradation in part as a regulator of SCF ubiquitin E3 ligases. The CSN is important for timely germination of seeds, but its molecular targets in this process is unclear. In this study, we demonstrate that the CSN regulates protein stabilities of two different targets from two antagonistic hormonal pathways, RGL2 of the GA pathway and ABI5 of the ABA pathway. Our genetic and transcriptome analyses showed that, although csn1-10 and csn5a-1 exhibit similar defects in timely germination, the mechanisms of how the mutations affect seed germination differ. Since RGL2 is known to be targeted by SCF during germination, the defect in the timely degradation of RGL2 in csn1-10 and csn5a-1 is consistent with the role of CSN as a regulator of the SCF. In addition, we show that CSN5A, but not CSN1, has an additional function in regulating ABI5, a downstream inhibitor of germination.