Caged phosphopeptides reveal a temporal role for 14-3-3 in G1 arrest and S-phase checkpoint function.
Nat Biotechnol
; 22(8): 993-1000, 2004 Aug.
Article
em En
| MEDLINE
| ID: mdl-15273693
ABSTRACT
Using classical genetics to study modular phosphopeptide-binding domains within a family of proteins that are functionally redundant is difficult when other members of the domain family compensate for the product of the knocked-out gene. Here we describe a chemical genetics approach that overcomes this limitation by using UV-activatable caged phosphopeptides. By incorporating a caged phosphoserine residue within a consensus motif, these reagents simultaneously and synchronously inactivate all phosphoserine/phosphothreonine-binding domain family members in a rapid and temporally regulated manner. We applied this approach to study the global function of 14-3-3 proteins in cell cycle control. Activation of the caged phosphopeptides by UV irradiation displaced endogenous proteins from 14-3-3-binding, causing premature cell cycle entry, release of G1 cells from interphase arrest and loss of the S-phase checkpoint after DNA damage, accompanied by high levels of cell death. This class of reagents will greatly facilitate molecular dissection of kinase-dependent signaling pathways when applied to other phosphopeptide-binding domains including SH2, Polo-box and tandem BRCT domains.
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Base de dados:
MEDLINE
Assunto principal:
Fosfopeptídeos
/
Fotoquímica
/
Osteossarcoma
/
Fase G1
/
Fase S
/
Proteínas 14-3-3
Limite:
Humans
Idioma:
En
Ano de publicação:
2004
Tipo de documento:
Article