10.1021/acs.jproteome.9b00373.s001
Gerald Dayebgadoh
Gerald
Dayebgadoh
Mihaela E. Sardiu
Mihaela E.
Sardiu
Laurence Florens
Laurence
Florens
Michael P. Washburn
Michael P.
Washburn
Biochemical Reduction
of the Topology of the Diverse
WDR76 Protein Interactome
American Chemical Society
2019
CCT
PARP
WDR 76
DNA-PK
WDR 76 interactome
WD 40 domain
gene expression regulation
protein interaction networks
study WDR 76
Diverse WDR 76 Protein Interactome
SIRT
WD 40
hub protein
protein quality control
DNA damage repair
WDR 76-based protein complexes
GAN
AP
WDR 76 interactions
2019-08-09 13:41:42
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Biochemical_Reduction_of_the_Topology_of_the_Diverse_WDR76_Protein_Interactome/9441713
A hub
protein in protein interaction networks will typically have
a large number of diverse interactions. Determining the core interactions
and the function of such a hub protein remains a significant challenge
in the study of networks. Proteins with WD40 repeats represent a large
class of proteins that can be hub proteins. WDR76 is a poorly characterized
WD40 repeat protein with possible involvement in DNA damage repair,
cell-cycle progression, apoptosis, gene expression regulation, and
protein quality control. WDR76 has a large and diverse interaction
network that has made its study challenging. Here we rigorously carry
out a series of affinity purification coupled to mass spectrometry
(AP–MS) analyses to map out the WDR76 interactome through different
biochemical conditions. We apply AP–MS analysis coupled to
size-exclusion chromatography to resolve WDR76-based protein complexes.
Furthermore, we also show that WDR76 interacts with the CCT complex
via its WD40 repeat domain and with DNA-PK–KU, PARP1, GAN,
SIRT1, and histones outside of the WD40 domain. An evaluation of the
stability of WDR76 interactions led to focused and streamlined reciprocal
analyses that validate the interactions with GAN and SIRT1. Overall,
the approaches used to study WDR76 would be valuable to study other
proteins containing WD40 repeat domains, which are conserved in a
large number of proteins in many organisms.