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The fission yeast COP9/signalosome is involved in cullin modification by ubiquitin-related Ned8p

Zhou, Chunshui and Seibert, Volker and Geyer, Rory and Rhee, Edward and Lyapina, Svetlana and Cope, Greg and Deshaies, Raymond J. and Wolf, Dieter A. (2001) The fission yeast COP9/signalosome is involved in cullin modification by ubiquitin-related Ned8p. BMC Biochemistry, 2 (7). pp. 1-11. ISSN 1471-2091. PMCID PMC37391.

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Background: The function of the fission yeast cullins Pcu1p and Pcu4p requires modification by the ubiquitin-related peptide Ned8p. A recent report by Lyapina et al. shows that the COP9/signalosome (CSN), a multifunctional eight subunit complex, regulates Ned8p modification of Pcu1p. Disruption of caa1/csn1, which encodes subunit 1 of the putative S. pombe CSN, results in accumulation of Pcu1p exclusively in the modified form. However, it remained unclear whether this reflects global control of all cullins by the entire CSN complex. Results: We demonstrate that multiple CSN subunits control Ned8p modification of Pcu3p, another fission yeast cullin, which, in complex with the RING domain protein Pip1p, forms a ubiquitin ligase that functions in cellular stress response. Pcu3p is modified by Ned8p on Lys 729 and accumulates exclusively in the neddylated form in cells lacking the CSN subunits 1, 3, 4, and 5. These CSN subunits co-elute with Pcu3p in gel filtration fractions corresponding to ~550 kDa and specifically bind both native and Ned8p-modified Pcu3p in vivo. While CSN does not influence the subcellular localization of Pcu3p, Pcu3p-associated in vitro ubiquitin ligase activity is stimulated in the absence of CSN. Conclusions: Taken together, our data suggest that CSN is a global regulator of Ned8p modification of multiple cullins and potentially other proteins involved in cellular regulation.

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Deshaies, Raymond J.0000-0002-3671-9354
Additional Information:Received: 3 May 2001; Accepted: 18 July 2001; Published: 18 July 2001. BMC Biochemistry 2001, 2:7. This article is available from: © 2001 Zhou et al; licensee BioMed Central Ltd. Verbatim copying and redistribution of this article are permitted in any medium for any non-commercial purpose, provided this notice is preserved along with the article's original URL. For commercial use, contact We thank F. McKeon for hybridoma lines, R. King for the human E1 plasmid, D. Griffiths for numerous technical suggestions, and C. Maki and Z. Yuan for critical reading of this manuscript. This work is supported by NIH grant GM59780 to DAW.
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Subject Keywords:CSN: COP9/signalosome; GST: Glutathione S-transferase; HA: hemagglutinin; SCF: SKP1/Cullin/F-box protein complex; UBC: Ubiquitin-conjugating enzyme
Issue or Number:7
PubMed Central ID:PMC37391
Record Number:CaltechAUTHORS:ZHObmcb01
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:266
Deposited By: Archive Administrator
Deposited On:13 May 2005
Last Modified:02 Oct 2019 22:31

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