Characterization and purification of the mammalian COP9 complex, a conserved nuclear regulator initially identified as a repressor of photomorphogenesis in higher plants.
The COP9 complex has been identified as a repressor of photomorphogenesis in Arabidopsis. Here we demonstrate that the COP9 complex is also present in mammals. Specific antibodies were generated against human counterparts of the Arabidopsis COP9 and COP11, the two known subunits of plant COP9 complex. Using these antibodies, we showed that indeed mammalian COP9 and COP11, also known as GPS1, could be coimmuno-precipitated using either of the two specific antibodies, definitively confirming that they are physically part of the same complex. Further, the mammalian COP9 and COP11/GPS1 were cofractionated in the same large molecular weight fractions of about 500 kDa and were absent from the monomeric fractions. The mammalian COP9 complex was present in all organs examined but abundances vary. Indirect immunofluorescence studies suggested that the mammalian COP9 complex is largely nuclear localized. Both conventional biochemical and affinity purifications of the COP9 complex from pig spleen indicated that the mammalian COP9 complex consists of eight distinct subunits. These findings indicate that mammals also have a COP9 complex with conserved molecular composition and biochemical and cellular properties similar to the higher plant counterpart.
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520-8104, USA. nwei@peaplant.biology.yale.edu
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D000818:Animals; D017360:Arabidopsis / Q000254:growth & development / Q000378:metabolism / Q000528:radiation effects; D000075686:COP9 Signalosome Complex; D006801:Humans; D046911:Macromolecular Substances; D008322:Mammals; D008970:Molecular Weight; D046912:Multiprotein Complexes; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010447:Peptide Hydrolases; D018462:Photobiology; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011487:Protein Conformation; D011506:Proteins
D046911:Macromolecular Substances; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D010940:Plant Proteins; D011506:Proteins; D010447:Peptide Hydrolases; D000075686:COP9 Signalosome Complex