Biochemical evidence for ubiquitin ligase activity of the Arabidopsis COP1 interacting protein 8 (CIP8)

Details

Serval ID
serval:BIB_65DB54704EA8
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Biochemical evidence for ubiquitin ligase activity of the Arabidopsis COP1 interacting protein 8 (CIP8)
Journal
Plant Journal
Author(s)
Hardtke  C. S., Okamoto  H., Stoop-Myer  C., Deng  X. W.
ISSN
0960-7412 (Print)
Publication state
Published
Issued date
05/2002
Volume
30
Number
4
Pages
385-94
Notes
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S. --- Old month value: May
Abstract
Arabidopsis COP1 is a negative regulator of photomorphogenesis, which targets HY5, a positive regulator of photomorphogenesis, for degradation via the proteasome pathway in the absence of light. COP1 and its interactive partner CIP8 both possess RING finger motifs, characteristic of some E3 ubiquitin ligases. Here we show that CIP8 promotes ubiquitin attachment to HY5 in E2-dependent fashion in vitro. CIP8 exhibits a strong interaction with the E2 enzyme AtUBC8 through its N-terminal domain. Phosphorylation of HY5 by casein kinase II requires the beta subunit 2, but does not affect HY5's susceptibility to ubiquitination. The RING domain of CIP8 is required but is not sufficient for ubiquitin ligase activity. Although the RING domain of CIP8 interacts with the RING domain of COP1, addition of recombinant COP1 fails to affect CIP8's ubiquitin ligase activity towards HY5 in vitro. However, recombinant COP1 can pull-down native CIP8 from the extract of dark-grown seedlings, but not from the extract of light-grown seedlings in a column-binding assay, implying a requirement for light-regulated modification in vivo. Our data suggest that CIP8 can form a minimal ubiquitin ligase in co-operation with the E2 enzyme AtUBC8. It is possible that the AtUBC8-CIP8 module might interact with COP1 in vivo, thereby participating in proteasome-mediated degradation of HY5.
Keywords
Arabidopsis/*enzymology/genetics Arabidopsis Proteins/*metabolism Basic-Leucine Zipper Transcription Factors Carrier Proteins/genetics/*metabolism Casein Kinase II Darkness Leucine Zippers/physiology Ligases/*metabolism Light Morphogenesis/radiation effects Nuclear Proteins/metabolism Phosphorylation Protein Binding Protein Interaction Mapping Protein-Serine-Threonine Kinases/metabolism Repressor Proteins/metabolism *Ubiquitin-Conjugating Enzymes Ubiquitin-Protein Ligases Ubiquitins/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 20:51
Last modification date
20/08/2019 15:21
Usage data