The Arabidopsis CSN5A and CSN5B subunits are present in distinct COP9 signalosome complexes, and mutations in their JAMM domains exhibit differential dominant negative effects on development.
The COP9 signalosome (CSN) is an evolutionarily conserved multisubunit protein complex involved in a variety of signaling and developmental processes through the regulation of protein ubiquitination and degradation. A known biochemical role attributed to CSN is a metalloprotease activity responsible for the derubylation of cullins, core components for several types of ubiquitin E3 ligases. The CSN's derubylation catalytic center resides in its subunit 5, which in Arabidopsis thaliana is encoded by two homologous genes, CSN5A and CSN5B. Here, we show that CSN5A and CSN5B subunits are assembled into distinct CSN complexes in vivo, which are present in drastically different abundances, with CSN(CSN5A) appearing to be the dominant one. Transgenic CSN5A and CSN5B proteins carrying a collection of single mutations in or surrounding the metalloprotease catalytic center are properly assembled into CSN complexes, but only mutations in CSN5A result in a pleiotropic dominant negative phenotype. The extent of phenotypic effects caused by mutations in CSN5A is reflected at the molecular level by impairment in Cullin1 derubylation. These results reveal that three key metal binding residues as well as two other amino acids outside the catalytic center play important roles in CSN derubylation activity. Taken together, our data provide physiological evidence on a positive role of CSN in the regulation of Arabidopsis SCF (for Skp1-Cullin-F-box) E3 ligases through RUB (for Related to Ubiquitin) deconjugation and highlight the unequal role that CSN(CSN5A) and CSN(CSN5B) play in controlling the cellular derubylation of cullins. The initial characterization of CSN5A and CSN5B insertion mutants further supports these findings and provides genetic evidence on their unequal role in plant development.
Gusmaroli
Giuliana
G
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520-8104, USA.
Feng
Suhua
S
Deng
Xing Wang
XW
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000075686:COP9 Signalosome Complex; D020134:Catalytic Domain / Q000235:genetics; D018797:Cell Cycle Proteins / Q000378:metabolism; D044842:Cullin Proteins / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D005799:Genes, Dominant; D008025:Ligases / Q000378:metabolism; D046911:Macromolecular Substances / Q000378:metabolism; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes; D010447:Peptide Hydrolases; D010641:Phenotype; D030821:Plants, Genetically Modified; D017354:Point Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D044843:SKP Cullin F-Box Protein Ligases / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology
D029681:Arabidopsis Proteins; C497728:COP9 signalosome subunit 5, Arabidopsis; D018797:Cell Cycle Proteins; C100318:Cullin 1; D044842:Cullin Proteins; D004269:DNA, Bacterial; D046911:Macromolecular Substances; D046912:Multiprotein Complexes; D021122:Protein Subunits; D011506:Proteins; C036483:T-DNA; D044843:SKP Cullin F-Box Protein Ligases; D010447:Peptide Hydrolases; D000075686:COP9 Signalosome Complex; D008025:Ligases; C487541:RCE1 protein, Arabidopsis