115 research outputs found
An Unbiased Evaluation of CK2 Inhibitors by Chemoproteomics: Characterization of Inhibitor Effects on CK2 and Identification of Novel Inhibitor Targets
Recently protein kinases have emerged as some of the most promising drug targets; and therefore, pharmaceutical strategies have been developed to inhibit kinases in the treatment of a variety of diseases. CK2 is a serine/threonine-protein kinase that has been implicated in a number of cellular processes, including maintenance of cell viability, protection of cells from apoptosis, and tumorigenesis. Elevated CK2 activity has been established in a number of cancers where it was shown to promote tumorigenesis via the regulation of the activity of various oncogenes and tumor suppressor proteins. Consequently the development of CK2 inhibitors has been ongoing in preclinical studies, resulting in the generation of a number of CK2-directed compounds. In the present study, an unbiased evaluation of CK2 inhibitors 4,5,6,7-tetrabromo-
The FNIP co-chaperones decelerate the Hsp90 chaperone cycle and enhance drug binding
The ability of Heat Shock Protein 90 (Hsp90) to hydrolyze ATP is essential for its chaperone function. The co-chaperone Aha1 stimulates Hsp90 ATPase activity, tailoring the chaperone function to specific "client" proteins. The intracellular signaling mechanisms directly regulating Aha1 association with Heat shock protein-90 (Hsp90) is an essential molecular chaperone in eukaryotes involved in maintaining the stability and activity of numerous signalling proteins, also known as clients. Hsp90 ATPase activity is essential for its chaperone function and it is regulated by co-chaperones. Here we show that the tumour suppressor FLCN is an Hsp90 client protein and its binding partners FNIP1/FNIP2 function as co-chaperones. FNIPs decelerate the chaperone cycle, facilitating FLCN interaction with Hsp90, consequently ensuring FLCN stability. FNIPs compete with the activating co-chaperone Aha1 for binding to Hsp90, thereby providing a reciprocal regulatory mechanism for chaperoning of client proteins. Lastly, downregulation of FNIPs desensitizes cancer cells to Hsp90 inhibitors, whereas FNIPs overexpression in renal tumours compared with adjacent normal tissues correlates with enhanced binding of Hsp90 to its inhibitors. Our findings suggest that FNIPs expression can potentially serve as a predictive indicator of tumour response to Hsp90 inhibitors
New Modularity of DAP-Kinases: Alternative Splicing of the DRP-1 Gene Produces a ZIPk-Like Isoform
DRP-1 and ZIPk are two members of the Death Associated Protein Ser/Thr Kinase
(DAP-kinase) family, which function in different settings of cell death
including autophagy. DAP kinases are very similar in their catalytic domains but
differ substantially in their extra-catalytic domains. This difference is
crucial for the significantly different modes of regulation and function among
DAP kinases. Here we report the identification of a novel alternatively spliced
kinase isoform of the DRP-1 gene, termed DRP-1β. The
alternative splicing event replaces the whole extra catalytic domain of DRP-1
with a single coding exon that is closely related to the sequence of the extra
catalytic domain of ZIPk. As a consequence, DRP-1β lacks the calmodulin
regulatory domain of DRP-1, and instead contains a leucine zipper-like motif
similar to the protein binding region of ZIPk. Several functional assays proved
that this new isoform retained the biochemical and cellular properties that are
common to DRP-1 and ZIPk, including myosin light chain phosphorylation, and
activation of membrane blebbing and autophagy. In addition, DRP-1β also
acquired binding to the ATF4 transcription factor, a feature characteristic of
ZIPk but not DRP-1. Thus, a splicing event of the DRP-1 produces a ZIPk like
isoform. DRP-1β is highly conserved in evolution, present in all known
vertebrate DRP-1 loci. We detected the corresponding mRNA and
protein in embryonic mouse brains and in human embryonic stem cells thus
confirming the in vivo utilization of this isoform. The
discovery of module conservation within the DAPk family members illustrates a
parsimonious way to increase the functional complexity within protein families.
It also provides crucial data for modeling the expansion and evolution of DAP
kinase proteins within vertebrates, suggesting that DRP-1 and ZIPk most likely
evolved from their ancient ancestor gene DAPk by two gene duplication events
that occurred close to the emergence of vertebrates
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