Document Detail

p53 Ubiquitination and Proteasomal Degradation.
MedLine Citation:
PMID:  23150437     Owner:  NLM     Status:  Publisher    
p53 levels and activity are controlled in large part through regulated ubiquitination and subsequent destruction by the 26S proteasome. Monoubiquitination of p53 is mediated primarily by the RING-finger E3 ubiquitin ligase MDM2 and impacts p53 activity through modulation of p53 localization and transcription activities. Recently, several E4 ubiquitin ligases (E4s) have been identified which serve to extend these monoubiquitin chains. The ubiquitin ligase activity of these factors toward p53, and their contribution to p53 degradation, can be studied using a variety of in vitro and in vivo methods and reagents which will be described in this chapter. These methods include in vivo ubiquitination of p53 using HA-ubiquitin or his-ubiquitin; the in vitro E3 ubiquitin ligase assay, in which ubiquitin reaction components (URC) are incubated with a purified E3 or E4 ligase; a one-step E4 assay, in which URC are incubated with a substrate, E3, and E4; and a two-step E4 assay in which p53 is monoubiquitinated in an E3 reaction, and subsequently purified and incubated with an E4. Finally, we will describe an in vitro degradation assay in which ubiquitinated p53 is incubated with purified 26S proteasomes. Together, these assays can be used to provide insight into the biochemical nature of p53 ubiquitination and degradation.
Ian M Love; Dingding Shi; Steven R Grossman
Related Documents :
15547757 - Immunohistochemical staining of metallothionein in canine mammary tumors: better surviv...
16524907 - Posttranslational, translational, and transcriptional responses to nitric oxide stress ...
18633117 - Activation of nf-e2-related factor-2 reverses biochemical dysfunction of endothelial ce...
17362897 - Downregulation of cuzn-superoxide dismutase contributes to beta-adrenergic receptor-med...
3780677 - Suppression of urokinase-type plasminogen activator mrna levels in human fibrosarcoma c...
15177117 - Expression of the third complement component (c3) and carboxypeptidase n small subunit ...
Publication Detail:
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  962     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  63-73     Citation Subset:  -    
Department of Internal Medicine, Virginia Commonwealth University, Richmond, VA, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  p53 and Cell Cycle Effects After DNA Damage.
Next Document:  Identification of p53 in Mitochondria.