TY - JOUR A1 - Hagen, Sven A1 - Mattay, Dinah A1 - Raeuber, Christina A1 - Mueller, Kristian M. A1 - Arndt, Katja Maren T1 - Characterization and inhibition of AF10-mediated interaction JF - Journal of peptide science N2 - The non-random chromosomal translocations t(10;11)(p13;q23) and t(10;11)(p13;q14-21) result in leukemogenic fusion proteins comprising the coiled coil domain of the transcription factor AF10 and the proteins MLL or CALM, respectively, and subsequently cause certain types of acute leukemia. The AF10 coiled-coil domain, which is crucial for the leukemogenic effect, has been shown to interact with GAS41, a protein previously identified as the product of an amplified gene in glioblastoma. Using sequential synthetic peptides, we mapped the potential AF10/GAS41 interaction site, which was subsequently be used as scaffold for a library targeting the AF10 coiled-coil domain. Using phage display, we selected a peptide that binds the AF10 coiled-coil domain with higher affinity than the respective coiled-coil region of wild-type GAS41, as demonstrated by phage ELISA, CD, and PCAs. Furthermore, we were able to successfully deploy the inhibitory peptide in a mammalian cell line to lower the expression of Hoxa genes that have been described to be overexpressed in these leukemias. This work dissects molecular determinants mediating AF10-directed interactions in leukemic fusions comprising the N-terminal parts of the proteins MLL or CALM and the C-terminal coiled-coil domain of AF10. Furthermore, it outlines the first steps in recognizing and blocking the leukemia-associated AF10 interaction in histiocytic lymphoma cells and therefore, may have significant implications in future diagnostics and therapeutics. Copyright (c) 2014 European Peptide Society and John Wiley & Sons, Ltd. KW - protein-protein interaction KW - protein design and selection KW - protein engineering KW - coiled coil KW - leucine zipper KW - AF10 Y1 - 2014 U6 - https://doi.org/10.1002/psc.2626 SN - 1075-2617 SN - 1099-1387 VL - 20 IS - 6 SP - 385 EP - 397 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Nietzsche, Madlen A1 - Schiessl, Ingrid A1 - Börnke, Frederik T1 - The complex becomes more complex: protein-protein interactions of SnRK1 with DUF581 family proteins provide a framework for cell and stimulus type-specific SnRK1 signaling in plants JF - Frontiers in plant science N2 - In plants, SNF1-related kinase (SnRK1) responds to the availability of carbohydrates as well as to environmental stresses by down-regulating ATP consuming biosynthetic processes, while stimulating energy-generating catabolic reactions through gene expression and post-transcriptional regulation. The functional SnRK1 complex is a heterotrimer where the catalytic alpha subunit associates with a regulatory beta subunit and an activating gamma subunit. Several different metabolites as well as the hormone abscisic acid (ABA) have been shown to modulate SnRK1 activity in a cell- and stimulus-type specific manner. It has been proposed that tissue- or stimulus-specific expression of adapter proteins mediating SnRK1 regulation can at least partly explain the differences observed in SnRK1 signaling. By using yeast two-hybrid and in planta bi-molecular fluorescence complementation assays we were able to demonstrate that proteins containing the domain of unknown function (DUF) 581 could interact with both isoforms of the SnRK1 alpha subunit (AKIN10/11) of Arabidopsis. A structure/function analysis suggests that the DUF581 is a generic SnRK1 interaction module and co-expression with DUF581 proteins in plant cells leads to reallocation of the kinase to specific regions within the nucleus. Yeast two-hybrid analyses suggest that SnRK1 and DUF581 proteins share common interaction partners inside the nucleus. The analysis of available microarray data implies that expression of the 19 members of the DUF581 encoding gene family in Arabidopsis is differentially regulated by hormones and environmental cues, indicating specialized functions of individual family members. We hypothesize that DUF581 proteins could act as mediators conferring tissue- and stimulus-type specific differences in SnRK1 regulation. KW - Arabidopsis KW - SnRK1 KW - DUF581 KW - protein-protein interaction KW - stress signaling KW - ABA Y1 - 2014 U6 - https://doi.org/10.3389/fpls.2014.00054 SN - 1664-462X VL - 5 PB - Frontiers Research Foundation CY - Lausanne ER -