Inhibitors of HDACs are used in cancer therapy based on the role HDACs play in transcription by regulating chromatin
compaction and non-histone proteins such as transcription factors. Profiling of HDAC expression is of interest in the functional proteomics analysis of cancer. Also, non-HDAC proteins may interact with HDAC inhibitor drugs and contribute to the drug mode of action. We here present a tool for the unbiased chemical proteomic profiling of proteins that specifically interact with SAHA. We designed and synthesized a trifunctional Capture Compound containing SAHA as selectivity and identified HDACs1, 2, 3 and 6, known and predicted HDAC interactors from human-derived HepG2 cell lysate, as well as a set of new potential non-HDAC targets of SAHA. One of these non-HDAC targets, isochorismatase PS-341 in vivo domain-containing protein 2 (ISOC2) is putative hydrolase associated with the negative regulation of the tumor-suppressor p16(INK4a). Dibutyryl-cAMP We demonstrated the direct and dose-dependent interaction of SAHA to the purified recombinant ISOC2 protein. Using SAHA Capture Compound mass spectrometry, we thus identified potential new SAHA target proteins
in an entirely unbiased chemical proteomics approach.”
“Signaling via the epidermal growth factor receptor (EGFR) pathway has emerged as one of the key mechanisms in the development of the central nervous system in Drosophila melanogaster. By contrast, little is known about the functions of EGFR signaling in the differentiated larval brain. Here, promoter-reporter lines of EGFR and its most prominent activating ligands, Spitz, Keren, and Vein, were used to identify the brain structures relevant for the EGFR pathway. Unexpectedly, promoter activity of all these pathway components was found in the mushroom bodies, which are known to be a higher brain center required for olfactory learning. We investigated the role of the EGFR pathway in this process by using different mutant larvae with reduced pan-neuronal EGFR signaling and those with reduced EGFR signaling
in mushroom bodies only. Expression of a dominant-negative form of EGFR as well as silencing of the ligands via RNA interference was applied and resulted in significantly impaired olfactory learning Bacterial neuraminidase performances. General defects in the ability to taste or smell as well as impaired EGFR signaling during embryonic development could be excluded as major reasons for this learning phenotype. In addition, targeted expression of a constitutively active form of the ligand Spitz also led to a significantly reduced learning ability. Thus, very low levels as well as very high levels of EGFR signaling are deleterious for olfactory learning and memory formation. We hypothesize that EGFR signaling in a certain range maintains a homeostatic situation in the mushroom bodies that is necessary for proper learning and memory.