Project description:To systematically investigate the regulations and functions of human WW-domain containing proteins, we conducted a proteomic analysis of 26 full-length WW-domain containing proteins and 17 WW-domains only, and identified their associated protein complexes in human HEK293T cells using tandem affinity purifications followed by LC-MSMS.
Project description:Primary microRNA (pri-miRNA) transcripts are processed by a protein complex called the Microprocessor comprised of the ribonuclease Drosha and its RNA binding partner DGCR8/Pasha. We sequenced small RNAs from animals containing a mutation in the WW domain of C. elegans pash-1. We found that these mutants have a modest but widespread reduction in miRNA levels when grown at 20˚C, which is further enhanced when grown at 25˚C. The results demonstrate a requirement for the WW domain in processing miRNAs in C. elegans.
Project description:A total of 12 WW domains from Saccharomyces cerevisiae were expressed and purified as fusion proteins to either GST or MBP. The fusion proteins were chemically biotinylated and applied to duplicate protein microarrays. Data processing revealed a total of 587 interactions between the domains and 207 proteins. Most of these interactions have not been previously observed. Keywords: Protein microarray analysis of yeast WW domains
Project description:Extracellular vesicles (EVs), spherical nano-sized lipid-bilayer vesicular structure, is produced from host cells, and widely studied in microbiology, neurobiology, and bioengineering as drug delivery. Here, we report the new vaccine platform using a novel type of EVs, WW domain activated extracellular vesicles (WAEV). Our proteomics result showed that WAEV was activated by the WW domain independently, and distinct from the existing exosome.
Project description:The multidomain protein BAG3 exerts pleiotropic oncogenic functions in many tumor entities including glioblastoma (GBM). Here we compared BAG3 protein-protein interactions in either adherently cultured or stem-like cultured U251 GBM cells. In line with BAG3´s putative role in stemness, identified interactors in sphere-cultured cells included different stem cell markers (SOX2, OLIG2, NES), while interactomes of adherent BAG3-proficient cells indicated a shift towards involvement of BAG3 in regulation of cilium assembly (ACTR3, ARL3). Applying a set of BAG3 deletion constructs we could demonstrate that none of the domains except the WW domain are required for suppression of cilia formation by full-length BAG3 in U251 and U343 cells. In line with the established regulation of the Hippo pathway by this domain, we could show that the WW mutant fails to rescue YAP1 nuclear translocation. BAG3 depletion reduced activation of a YAP1/AURKA signaling pathway and induction of PLK1. Collectively, our findings point to a complex interaction network of BAG3 with several pathways regulating cilia homeostasis, involving processes related to ciliogenesis and cilium degradation.
