title: Assessment of HDACi-Induced Protein Cleavage by Caspases.
abstract: Aberrant histone deacetylase (HDAC) activity often correlates with neoplastic transformation and inhibition of HDACs by small molecules has emerged as a promising strategy to treat hematological malignancies in particular. Treatment with HDAC inhibitors (HDACis) often prompts tumor cells to undergo apoptosis, thereby causing a caspase-dependent cleavage of target proteins. An unexpectedly large number of proteins are in vivo caspase substrates and defining caspase-mediated substrate specificity is a major challenge. In this chapter we demonstrate that the hematopoietic transcription factor PU.1 becomes cleaved after treatment of acute myeloid leukemia (AML) cells with the HDACis LBH589 (panobinostat) or MS-275 (entinostat). To define caspase specificity for PU.1, an in vitro caspase assay including caspases 1-10 with in vitro-translated PU.1 is described in detail.
authors.affiliation: Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Treude
authors.affiliation: Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Gladbach
authors.affiliation: Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Plaster
authors.affiliation: Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany. jhartkamp@ukaachen.de.
authors.forename: Jörg
authors.initials: J
authors.lastname: Hartkamp
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D053181:Caspase 8 / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004789:Enzyme Activation; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D018922:HL-60 Cells; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D020014:K562 Cells; D000077767:Panobinostat; D059748:Proteolysis; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D013379:Substrate Specificity; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D053181:Caspase 8; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006877:Hydroxamic Acids; D007211:Indoles; D000077767:Panobinostat; D011518:Proto-Oncogene Proteins; D011725:Pyridines; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C118739:entinostat; C505530:CASP8 protein, human
keywords: Caspase-8; HDACi; In vitro translation; LBH589; MS-275; PU.1