Dear all,

On December 18, we welcome Ulrich Stelzl, from the University of Graz and BioTechMed-Graz, Austria, for an external seminar entitled

"Functional characterization of protein networks through deep mutational protein-interaction perturbation scanning"

at the Hexagone, at 10am

Ulrich Stelzl is a pioneer of human interactomics. 

His research group is focusing on the analysis of molecular interaction networks to understand the dynamics of molecular networks underlying cellular processes related to human diseases. They use a combination of experimental functional genomics techniques, such as high throughput yeast two-hybrid screening, biochemical, cell biological and computational methods. They currently focus on the systematic analyses of the functional impact of genetic variation and post-translational protein modification on protein-protein interaction using deep scanning mutagenesis approaches to bridge nucleotide resolution genomics and protein resolution proteomics.

Abstract

Deep mutational scanning approaches that assess the functional effects of thousands of missense variants of a protein is about to become a prime tool in functional genomics. Yeast based protein-protein interaction assays, that enrich interacting protein variants from non-interacting variants, such as yeast protein fragment complementation or yeast two-hybrid analysis, were used to scan several key protein interactions or small PPI networks. We developed an amino acid resolution protein-protein interaction perturbation scanning strategy to characterize protein function on an “edgetic” level. We use comprehensive single amino acid protein variant libraries and reverse yeast two-hybrid growth selection for efficient enrichment of non-interacting mutant protein versions, integrate our perturbation variant profiles with genetic and structural data and perform functional validation of key protein variants.

Assaying small protein networks, such as a target protein with a set of interaction partners, provides insight into protein function, cellular pathways or processes. We report deep scanning mutagenesis interaction perturbation screening results centered on tumor suppressor NF2 that reveal new clues to the conformation-dependent regulation of NF2 tumor suppressor activity. Deep mutational scanning provided amino acid resolution interaction surface information for ATGL`s most important four regulatory interaction partners involved inintracellular lipolysis. Specifically, we aim to elucidate and characterize “switch” mutations which are key in mediating interactions specifically to one of the partners with no effect on the others. Switch mutations are most selective prime tools for functional perturbation in varying biological context.

Key publications

Kohlmayr JM, Grabner GF, Nusser A, Höll A, Manojlović V, Halwachs B, Masser S, Jany-Luig E, Engelke E, Zimmermann R, Stelzl U (2024); Mutational scanning pinpoints distinct binding sites of key ATGL regulators in lipolysis; Nat Commun 15, 2516; doi: 10.1038/s41467-024-46937-x 

Jehle S, Kunowska N, Benlasfer N, Woodsmith J, Weber G, Wahl MC, Stelzl U (2022); A human kinase yeast array for the identification of kinases modulating phosphorylation-dependent protein-protein interactions; Mol Syst Biol 18, e10820; doi: 10.15252/msb.202110820 

Woodsmith J, Apelt L, Casado-Medrano V, Özkan Z, Timmermann B, Stelzl U (2017). Protein interaction perturbation profiling at amino acid resolution. Nat Methods 14, 1548-7091; doi: 10.1038/nmeth.4464 

Grossmann A, Benlasfer N, Birth P, Hegele A, Wachsmuth F, Apelt L, Stelzl U (2015). Phospho-tyrosine dependent protein-protein interaction network. Mol Syst Biol 11, 794; doi: 10.15252/msb.20145968

Hegele A, Kamburov A, Grossmann A, Sourlis C, Wowro S, Weimann M, Will CL, Pena V, Lührmann R, Stelzl U (2012). Dynamic protein-protein interaction wiring of the human spliceosome, Mol Cell 45, 567-580; doi: 10.1016/j.molcel.2011.12.034

Stelzl U, Worm U, Lalowski M, Haenig C, Brembeck FH, Goehler H, Stroedicke M, Zenkner M, Schoenherr A, Koeppen S, Timm J, Mintzlaff S, Abraham C, Bock N, Kietzmann S, Goedde A, Toksöz E, Droege A, Krobitsch S, Korn B, Birchmeier W, Lehrach H, Wanker EE (2005). A Human Protein-Protein Interaction Network: A Resource for Annotating the Proteome. Cell 122, 957-968; doi: 10.1016/j.cell.2005.08.029