Mutation of the Etr1 Ethylene Receptor. Ii: Reversion to Ethylene-sensitivity3 Is a Regulator of Seedling Growth

Title of Document: SUPPRESSORS OF etr1-2: I. etr1-11 IS A LOSS-OF-FUNCTION MUTATION OF THE ETR1 ETHYLENE RECEPTOR. II: REVERSION TO ETHYLENE-SENSITIVITY3 IS A REGULATOR OF SEEDLING GROWTH Christopher Alan McClellan, Doctor of Philosophy, 2009 Directed By: Dr. Caren Chang, Department of Cell Biology and Molecular Genetics The plant hormone ethylene is an important regulator of plant growth and development, including senescence, abcission, fruit ripening, and responses to biotic and abiotic stresses. To find new members of the ethylene signaling pathway, a genetic screen for suppressors of the ethylene-insensitive mutant etr1-2 was performed. One mutant identified in this screen, etr1-11, is an intragenic mutation within ETR1. etr1-11 is a unique missense mutation that appears to eliminate ETR12 signaling. Mutant analysis further revealed that etr1-11 is a partial loss-of-function allele. The rte3 (reversion to ethylene sensitivity3) mutant was another mutant isolated in a genetic screen for suppressors of etr1-2. After testing other ethylene responses, such as leaf senescence, and performing epistasis analysis with other ethylene signaling mutants, it was determined that RTE3 is unlikely to play a direct role in the ethylene signaling pathway. Instead, RTE3 appears to be responsible for promoting hypocotyl elongation in etiolated seedlings in the ethylene triple response assay. The RTE3 gene was identified by positional cloning, and is predicted to encode a protein with an annotated SAC3/GANP domain. SAC3/GANP domains are present in proteins that participate in large multi-peptide complexes, such as the 26S proteasome regulatory subunit and the eIF3 translation initiation complex. Similarities in protein composition between these two complexes and the COP9 signalosome (CSN) suggest that a SAC3/GANP domain-containing protein may interact with members of the CSN. Interestingly, yeast two-hybrid analysis reveals that RTE3 interacts with EER5 and EIN2, proteins that have been shown to interact with members of the CSN. In addition, rte3-1 ein2-1 seedlings show a synthetic phenotype of delayed growth. Protein localization using a GFP tag reveals that RTE3 and EER5 both localize to the nucleus. These interactions suggest that RTE3, EER5, EIN2, and the CSN form a protein complex that regulates seedling growth. SUPPRESSORS OF etr1-2: I. etr1-11 IS A LOSS-OF-FUNCTION MUTATION OF THE ETR1 ETHYLENE RECEPTOR. II: REVERSION TO ETHYLENE-SENSITIVITY3 IS A REGULATOR OF SEEDLING GROWTH