TY - JOUR A1 - Radu, Laura A1 - Schoenwetter, Elisabeth A1 - Braun, Cathy A1 - Marcoux, Julien A1 - Koelmel, Wolfgang A1 - Schmitt, Dominik R. A1 - Kuper, Jochen A1 - Cianférani, Sarah A1 - Egly, Jean M. A1 - Poterszman, Arnaud A1 - Kisker, Caroline T1 - The intricate network between the p34 and p44 subunits is central to the activity of the transcription/DNA repair factor TFIIH JF - Nucleic Acids Research N2 - The general transcription factor IIH (TFIIH) is a multi-protein complex and its 10 subunits are engaged in an intricate protein–protein interaction network critical for the regulation of its transcription and DNA repair activities that are so far little understood on a molecular level. In this study, we focused on the p44 and the p34 subunits, which are central for the structural integrity of core-TFIIH. We solved crystal structures of a complex formed by the p34 N-terminal vWA and p44 C-terminal zinc binding domains from Chaetomium thermophilum and from Homo sapiens. Intriguingly, our functional analyses clearly revealed the presence of a second interface located in the C-terminal zinc binding region of p34, which can rescue a disrupted interaction between the p34 vWA and the p44 RING domain. In addition, we demonstrate that the C-terminal zinc binding domain of p34 assumes a central role with respect to the stability and function of TFIIH. Our data reveal a redundant interaction network within core-TFIIH, which may serve to minimize the susceptibility to mutational impairment. This provides first insights why so far no mutations in the p34 or p44 TFIIH-core subunits have been identified that would lead to the hallmark nucleotide excision repair syndromes xeroderma pigmentosum or trichothiodystrophy. KW - general transcription factor IIH (TFIIH) KW - DNA repair KW - protein–protein interaction KW - p44 KW - p34 Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170173 VL - 45 IS - 18 ER - TY - JOUR A1 - Kaiser, Sebastian A1 - Sauer, Florian A1 - Kisker, Caroline T1 - The structural and functional characterization of human RecQ4 reveals insights into its helicase mechanism JF - Nature Communications N2 - RecQ4 is a member of the RecQ helicase family, an evolutionarily conserved class of enzymes, dedicated to preserving genomic integrity by operating in telomere maintenance, DNA repair and replication. While reduced RecQ4 activity is associated with cancer predisposition and premature aging, RecQ4 upregulation is related to carcinogenesis and metastasis. Within the RecQ family, RecQ4 assumes an exceptional position, lacking several characteristic RecQ domains. Here we present the crystal structure of human RecQ4, encompassing the conserved ATPase core and a novel C-terminal domain that lacks resemblance to the RQC domain observed in other RecQ helicases. The new domain features a zinc-binding site and two distinct types of winged-helix domains, which are not involved in canonical DNA binding or helicase activity. Based on our structural and functional analysis, we propose that RecQ4 exerts a helicase mechanism, which may be more closely related to bacterial RecQ helicases than to its human family members. KW - x-ray crystallography KW - enzymes KW - RecQ4 KW - humans Y1 - 2017 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170769 VL - 8 IS - 15907 ER - TY - JOUR A1 - Petruseva, Irina A1 - Naumenko, Natalia A1 - Kuper, Jochen A1 - Anarbaev, Rashid A1 - Kappenberger, Jeannette A1 - Kisker, Caroline A1 - Lavrik, Olga T1 - The Interaction Efficiency of XPD-p44 With Bulky DNA Damages Depends on the Structure of the Damage JF - Frontiers in Cell and Developmental Biology N2 - The successful elimination of bulky DNA damages via the nucleotide excision repair (NER) system is largely determined by the damage recognition step. This step consists of primary recognition and verification of the damage. The TFIIH helicase XPD plays a key role in the verification step during NER. To date, the mechanism of damage verification is not sufficiently understood and requires further detailed research. This study is a systematic investigation of the interaction of ctXPD (Chaetomium thermophilum) as well as ctXPD-ctp44 with model DNAs, which contain structurally different bulky lesions with previously estimated NER repair efficiencies. We have used ATPase and DNA binding studies to assess the interaction of ctXPD with damaged DNA. The result of the analysis of ctXPD-ctp44 binding to DNA containing fluorescent and photoactivatable lesions demonstrates the relationship between the affinity of XPD for DNAs containing bulky damages and the ability of the NER system to eliminate the damage. Photo-cross-linking of ctXPD with DNA probes containing repairable and unrepairable photoactivatable damages reveals differences in the DNA interaction efficiency in the presence and absence of ctp44. In general, the results obtained indicate the ability of ctXPD-ctp44 to interact with a damage and suggest a significant role for ctp44 subunit in the verification process. KW - nucleotide excision repair KW - XPD helicase KW - DNA damage KW - protein-DNA interaction KW - bulky damages recognition KW - photo-cross-linking Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-231806 SN - 2296-634X VL - 9 ER - TY - JOUR A1 - Peissert, Stefan A1 - Sauer, Florian A1 - Grabarczyk, Daniel B. A1 - Braun, Cathy A1 - Sander, Gudrun A1 - Poterszman, Arnaud A1 - Egly, Jean-Marc A1 - Kuper, Jochen A1 - Kisker, Caroline T1 - In TFIIH the Arch domain of XPD is mechanistically essential for transcription and DNA repair JF - Nature Communications N2 - The XPD helicase is a central component of the general transcription factor TFIIH which plays major roles in transcription and nucleotide excision repair (NER). Here we present the high-resolution crystal structure of the Arch domain of XPD with its interaction partner MAT1, a central component of the CDK activating kinase complex. The analysis of the interface led to the identification of amino acid residues that are crucial for the MAT1-XPD interaction. More importantly, mutagenesis of the Arch domain revealed that these residues are essential for the regulation of (i) NER activity by either impairing XPD helicase activity or the interaction of XPD with XPG; (ii) the phosphorylation of the RNA polymerase II and RNA synthesis. Our results reveal how MAT1 shields these functionally important residues thereby providing insights into how XPD is regulated by MAT1 and defining the Arch domain as a major mechanistic player within the XPD scaffold. KW - nucleotide excision repair KW - nuclear receptors KW - helicase KW - transactivation KW - fluorescence KW - recognition KW - subunit KW - binding KW - sulfur KW - kinease Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-229857 VL - 11 IS - 1 ER -