Structural Biology of Transcriptional Regulators in Disease
Title: Structural Biology of Transcriptional Regulators in Disease
DNr: SNIC 2014/1-292
Project Type: SNIC Medium Compute
Principal Investigator: Maria Sunnerhagen <marsu@ifm.liu.se>
Affiliation: Linköpings universitet
Duration: 2014-09-29 – 2015-10-01
Classification: 10601 10603 10602
Keywords:

Abstract

SUMMARY In my research group, we evaluate the structure and dynamics of transcriptional regulators and their interactions with DNA, with each other, with the translational machinery, with chromatin regulators, and with systems that govern protein stability such as ubiquitination and phosphorylation. Our main objectives of study are 1) the oncogene c-Myc, a highly attractive target for cancer therapeutics where we investigate both its internal structural and dynamic properties and its interactions with regulatory proteins, 2) the E3 ligase Ro52, which regulates interferone levels in cells and thereby affects gene expresion, and finally 3) MexR, a direct regulator of gene expression in Pseudomonas Aeruginosa and a target in cystic fibrosis. In collaboration with prof Mikael Sigvardsson, HU/LiU, we are studying the transcriptional regulator Ebf1, which is essential for the differentiation of B-cells and thereby related to leukemia. Here, we combine the use of structural biology methods in my group with proteomics, bioinformatics and cellular biology, also in international collaboration with U Toronto. PROGRESS Our previous SNIC project has resulted in one publication and two manuscripts currently in submission: A. High-resolution structure of TBP with TAF1 reveals anchoring patterns in transcriptional regulation. Anandapadamanaban M, Andresen C, Helander S, Ohyama Y, Siponen MI, Lundström P, Kokubo T, Ikura M, Moche M, & Sunnerhagen M*. Nature Structural & Molecular Biology 20, 1008–1014 (2013) B. Pre-anchoring of Pin1 to unphosphorylated c-Myc in a dynamic complex affects c-Myc activity. Sara Helander, Meri Montecchio, Robert Pilstål, Yulong Su, Jacob Kuruvilla, Malin Johansson, Javed Mohammed, Susana Cristobal, Patrik Lundström, Rosalie Sears, Björn Wallner & Maria Sunnerhagen* (Nature Structure Molecular Biology 2014, submitted) C. A MexR antibiotics resistance mutation employs conformational selection to restrict access to DNA-binding states. Madhanagopal Anandapadamanaban, Robert Pilstål, Javed Muhammed, Cecilia Andrésen, Björn Wallner* and Maria Sunnerhagen*. (PNAS, pending submission). In addition to this, we are currently wrapping up the structure determination of Ro52 complexes with E2 conjugating enzymes UbcH6 and UbcH5, in the presence and absence of ubiquitin (Anandapadamanaban et al., 2014 manuscript in preparation), and we are refining the structure of a novel Myc-TBP complex of utmost importance for the biology of cancer (Montecchio, Wallenhammar et al., 2014 ongoing). UPCOMING Our main task during the upcoming year will be to further characterize dynamic Myc complexes by means of a combined use of NMR, biophysical and computational resources. We have preliminary data for a range of complexes which now await refinement. Furthermore, our joint translational approach to the proteomic study of Ebf1 interactions using structural, computational and biological methods is already using significant compute resources at Kappa and we desperately need more computational power to address these complex data. Finally, we hope during this upcoming year to be able to resolve in more detail the structural mechanism of ubiquitination by TRIM21, which holds key to how proteins are degraded in the cell. In all these respects, NSC and Triolith is a critical asset for us to achieve our scientific goals.