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Vol. 13, Issue 2, 646-655, February 2002

CSN1 N-Terminal-dependent Activity Is Required for Arabidopsis Development But Not for Rub1/Nedd8 Deconjugation of Cullins: A Structure-Function Study of CSN1 Subunit of COP9 Signalosome

Xiping Wang,^* Dingming Kang,^* Suhua Feng, Giovanna Serino, Claus Schwechheimer,^§ and Ning Wei

Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520

The COP9 signalosome (CSN) is a multifunctional protein complex essential for arabidopsis development. One of its functions is to promote Rub1/Nedd8 deconjugation from the cullin subunit of the Skp1-cullin-F-box ubiquitin ligase. Little is known about the specific role of its eight subunits in deneddylation or any of the physiological functions of CSN. In the absence of CSN1 (the fus6 mutant), arabidopsis CSN complex cannot assemble, which destabilizes multiple CSN subunits and contributes, together with the loss of CSN1, to the phenotype of fus6. To distinguish CSN1-specific functions, we attempted to rescue the complex formation with deletion or point-mutation forms of CSN1 expressed as transgenes in fus6. We show that the central domain of CSN1 is critical for complex assembly, whereas the C-terminal domain has a supporting role. By expressing the C231 fragment, which contains the structural information but lacks the presumed functional domain located at the N terminus, we have rescued the complex formation and restored the Rub1/Nedd8 deconjugation activity on cullins (fus6/C231). Nonetheless, fus6/C231 exhibits pleiotropic phenotype, including photomorphogenic defects and growth arrest at seedling stage. We conclude that CSN1 N-terminal domain is not required for the Rub1/Nedd8 deconjugation activity of cullins, but contributes to a significant aspect of CSN functions that are essential for plant development.

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^* These authors are co-first authors.

Present addresses: Fudan University, Department of Biochemistry, Shanghai 200433, China; Peking-Yale Joint Center for Plant Molecular Genetics and Agribiotechnology, Peking University, Beijing 100871, China; ^§ZMBP-Center for Plant Molecular Biology, Universität Tuebingen, Tuebingen, Germany.

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Molecular Biology of the Cell
Vol. 13, 646-655, February 2002
Copyright © 2002 by The American Society for Cell Biology

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