TY - JOUR
T1 - Synthesis of isomeric phosphoubiquitin chains reveals that phosphorylation controls deubiquitinase activity and specificity
AU - Huguenin-Dezot, Nicolas
AU - De Cesare, Virginia
AU - Peltier, Julien
AU - Knebel, Axel
AU - Kristariyanto, Yosua Adi
AU - Rogerson, Daniel T.
AU - Kulathu, Yogesh
AU - Trost, Matthias
AU - Chin, Jason W.
PY - 2016/7/26
Y1 - 2016/7/26
N2 - Ubiquitin is post-translationally modified by phosphorylation at several sites, but the consequences of these modifications are largely unknown. Here we synthesize multi-milligram quantities of ubiquitin phosphorylated at serine 20, serine 57 and serine 65 via genetic code expansion. We use these phosphoubiquitins for the enzymatic assembly of twenty isomeric phospho-ubiquitin dimers, with different sites of isopeptide linkage and/or phosphorylation. We discover that phosphorylation of serine 20 on ubiquitin converts UBE3C from a dual specificity E3 ligase into a ligase that primarily synthesizes K48 chains. We profile the activity of 31 deubiquitinases on the isomeric phosphoubiquitin dimers in 837 reactions, and discover that phosphorylation at distinct sites in ubiquitin can activate or repress cleavage of a particular linkage by deubiquitinases, and that phosphorylation at a single site in ubiquitin can control the specificity of deubiquitinases for distinct ubiquitin linkages.
AB - Ubiquitin is post-translationally modified by phosphorylation at several sites, but the consequences of these modifications are largely unknown. Here we synthesize multi-milligram quantities of ubiquitin phosphorylated at serine 20, serine 57 and serine 65 via genetic code expansion. We use these phosphoubiquitins for the enzymatic assembly of twenty isomeric phospho-ubiquitin dimers, with different sites of isopeptide linkage and/or phosphorylation. We discover that phosphorylation of serine 20 on ubiquitin converts UBE3C from a dual specificity E3 ligase into a ligase that primarily synthesizes K48 chains. We profile the activity of 31 deubiquitinases on the isomeric phosphoubiquitin dimers in 837 reactions, and discover that phosphorylation at distinct sites in ubiquitin can activate or repress cleavage of a particular linkage by deubiquitinases, and that phosphorylation at a single site in ubiquitin can control the specificity of deubiquitinases for distinct ubiquitin linkages.
U2 - 10.1016/j.celrep.2016.06.064
DO - 10.1016/j.celrep.2016.06.064
M3 - Article
C2 - 27425610
VL - 16
SP - 1180
EP - 1193
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 4
ER -