The functional and molecular architecture of core TFIIH
Final Report Abstract
In this this project we could successfully develop a novel low cost phasing protocol for macromolecular x-ray crystallography that can be easily implemented in many different laboratory setups. We have solved crystal structures of p52, p52/p8 and the PH domain p62. In addition, we could boost the capabilities to investigate higher order molecular complexes using single particle cryo EM, providing an excellent basis for further investigations on core TFIIH complexes in the presence of substrates and additional NER factors. We successfully deciphered the activity of the XPB enzyme. We utilized our crystallographic data of p52/p8 to investigate p52 mediated XPB activation. We have shown that the XPB ATPase can be activated by p52/p8 and DNA. However, when both components are present at the same time p52/p8 dominates and acts as a speed limiter even in the context of core TFIIH. Since the ATPase activity of XPB is vital for the essential processes of transcription and NER our results led to significant new insights on XPB activation and regulation within TFIIH using our unique modular assembly approach, thus redefining the regulatory network of XPB. Furthermore, we could show that despite full ATPase activity, XPB requires complete core TFIIH to act as a double stranded DNA translocase. In summary, we were able to examine XPB activation within core TFIIH at an unprecedented level that has led to an understanding of how XPB is implemented and regulated in the TFIIH machinery.
Publications
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How to limit the speed of a motor: the intricate regulation of the XPB ATPase and translocase in TFIIH. Nucleic Acids Research, 48(21), 12282-12296.
Kappenberger, Jeannette; Koelmel, Wolfgang; Schoenwetter, Elisabeth; Scheuer, Tobias; Woerner, Julia; Kuper, Jochen & Kisker, Caroline
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Cesium based phasing of macromolecules: a general easy to use approach for solving the phase problem. Scientific Reports, 11(1).
Koelmel, Wolfgang; Kuper, Jochen & Kisker, Caroline
