Caged Phosphopeptides Reveal a Temproal Role for 14-3-3 in G1 Arrest and S-phase Checkpoint Function  

Oleh:

Nguyen, Anhco ; Rothman, Debroah M. ; Stehn, Justine ; Imperiali, Barbara ; Yaffe, Michael B.

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 22 no. 8 (Agu. 2004), page 993-1000.
Topik: GENETICS; caged phosphopeptides; checkpoint function

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.1 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

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 phosposerine residue within a consensus motif, these reagents simultaneously and synchronously inactivate all phosphoserine / phospgothreonine 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 damafe, 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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