10 research outputs found
HOP_scores
Tab-delimited text file with MADL and z-scores for all approx. 1800 compounds tested in this stud
HIP_scores
Tab-delimited text file with MADL and z-scores for all approx. 1800 compounds tested in this stud
HIP_scores-benomyl
Tab delimited text file with the data of all performed Benomyl experiment
HOP_scores-benomyl
Tab delimited text file with the data of all performed Benomyl experiment
Evidence for a Functionally Relevant Rocaglamide Binding Site on the eIF4A–RNA Complex
Translation
initiation is an emerging target in oncology and neurobiology indications.
Naturally derived and synthetic rocaglamide scaffolds have been used
to interrogate this pathway; however, there is uncertainty regarding
their precise mechanism(s) of action. We exploited the genetic tractability
of yeast to define the primary effect of both a natural and a synthetic
rocaglamide in a cellular context and characterized the molecular
target using biochemical studies and <i>in silico</i> modeling.
Chemogenomic profiling and mutagenesis in yeast identified the eIF
(eukaryotic Initiation Factor) 4A helicase homologue as the primary
molecular target of rocaglamides and defined a discrete set of residues
near the RNA binding motif that confer resistance to both compounds.
Three of the eIF4A mutations were characterized regarding their functional
consequences on activity and response to rocaglamide inhibition. These
data support a model whereby rocaglamides stabilize an eIF4A-RNA interaction
to either alter the level and/or impair the activity of the eIF4F
complex. Furthermore, <i>in silico</i> modeling supports
the annotation of a binding pocket delineated by the RNA substrate
and the residues identified from our mutagenesis screen. As expected
from the high degree of conservation of the eukaryotic translation
pathway, these observations are consistent with previous observations
in mammalian model systems. Importantly, we demonstrate that the chemically
distinct silvestrol and synthetic rocaglamides share a common mechanism
of action, which will be critical for optimization of physiologically
stable derivatives. Finally, these data confirm the value of the rocaglamide
scaffold for exploring the impact of translational modulation on disease