GYF domain mediated protein: protein interactions
Zusammenfassung der Projektergebnisse
GYF domain containing proteins represent a small yet ubiquitously expressed family that is involved in mRNA surveillance. All of the family members contain a proline-rich sequence recognition domain termed GYF after a tripeptide that is part of a conserved set of mostly aromatic amino acids that form the binding pocket for the ligand. Two major subfamilies of GYF domains can be distinguished based on sequence homology and structural insights. The so-called CD2BP2 family comprises mostly nuclear proteins with a preference for peptides containing the motif PPGW while the larger Smy2 family binds best to PPGΦ motifs with Φ representing a hydrophobic or aromatic residue except for tryptophane. We have analysed the structure and interactome of certain SMY2 family members of GYF proteins from S. cerevisiae and H. sapiens and could assign a role in mRNA processing, transport and translational repression, in accordance with data from other laboratories such as the interaction of SMYII with COPII vesicular proteins. For CD2BP2-GYF we have analysed rationally designed peptidomimetic inhibitors in collaboration with project 3 of the research training group. A major focus of the work performed during this funding period was the creation and analysis of CD2BP2 knock-out mice. Constitutive deletion of the gene results in embryonic lethality at day 10.5 d.p.c. most probably resulting from mesodermal- derived tissue. Specific deletion in glomerular podocytes manifests in proteinuria and effacement, ultimately leading to kidney failure. At the molecular level we observe large changes in the alternative-splicing pattern of CD2BP2 knockout cells in either bone marrow-derived macrophages or podocytes. Most changes of transcript usage are cell-type specific while alternative splicing of exon 6 of VEGFA is seen in both experimental settings. Recruitment of the phosphatase PP1 to the maturing spliceosome is the proposed mechanism for the observed changes in alternative-splicing.
Projektbezogene Publikationen (Auswahl)
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(2010). Addressing protein protein interactions by small molecules: A designed Pro-Pro dipeptide mimic with a PPII helix conformation as a module for the synthesis of PRD-binding ligands. Angewandte Chemie, Int. Ed., 49, 7111-7115
Zaminer, J., Brockmann, C., Huy, P., Opitz, R., Reuter, C., Beyermann, M., Freund, C., Müller, M., Oschkinat, H., Kühne, R., and Schmalz, H.-G.
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(2010). Conserved β-hairpin recognition by the GYF domains of SMY2 and GIGYF2 in mRNA surveillance and vesicular transport complexes. Structure, 18, 944-954
Ash, M.-R., Fälber, K., Kosslick, D., Albert, G.I., Roske, Y., Kofler, M., Schümann, M., Krause, E. & Freund, C.
