Wahl, Markus C.

[["dc.bibliographiccitation.firstpage","531"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Journal of Molecular Biology"],["dc.bibliographiccitation.lastpage","541"],["dc.bibliographiccitation.volume","385"],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Luehrmann, Reinhard"],["dc.contributor.author","Wahl, Markus C."],["dc.date.accessioned","2018-11-07T08:33:29Z"],["dc.date.available","2018-11-07T08:33:29Z"],["dc.date.issued","2009"],["dc.description.abstract","The precursor mRNA retention and splicing (RES) complex mediates nuclear retention and enhances splicing of precursor mRNAs. The RES complex from yeast comprises three proteins, Snu17p, Bud13p and Pml1p. Snu17p acts as a central platform that concomitantly binds the Bud13p and Pml1p subunits via short peptide epitopes. As a step to decipher the molecular architecture of the RES complex, we have determined crystal structures of full-length Pml1p and N-terminally truncated Pml1p. The first 50 residues of full-length Pml1p, encompassing the Snu17p-binding region, are disordered, showing that Pml1p binds to Snu17p via an intrinsically unstructured region. The remainder of Pml1p folds as a forkhead-associated (FHA) domain, which is expanded by a number of noncanonical elements compared with known FHA domains from other proteins. An atypical N-terminal appendix runs across one beta-sheet and thereby stabilizes the domain as shown by deletion experiments. FHA domains are thought to constitute phosphopeptide-binding elements. Consistently, a sulfate ion was found at the putative phosphopeptide-binding loops of full-length Pml1p. The N-terminally truncated version of the protein lacked a similar phosphopeptide mimic but retained an almost identical structure. A long loop neighboring the putative phosphopeptide-binding site was disordered in both structures. Comparison with other FHA domain proteins suggests that this loop adopts a defined conformation upon ligand binding and thereby confers ligand specificity. Our results show that in the RES complex, an FHA domain of Pml1p is flexibly tethered via an unstructured N-terminal region to Snu17p. (C) 2008 Elsevier Ltd. All rights reserved."],["dc.description.sponsorship","Max Planck Society"],["dc.identifier.doi","10.1016/j.jmb.2008.10.087"],["dc.identifier.isi","000262916900016"],["dc.identifier.pmid","19010333"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/17588"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Academic Press Ltd Elsevier Science Ltd"],["dc.relation.issn","0022-2836"],["dc.title","Crystal Structure of the Pml1p Subunit of the Yeast Precursor mRNA Retention and Splicing Complex"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","13005"],["dc.bibliographiccitation.issue","32"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","13009"],["dc.bibliographiccitation.volume","104"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:49:26Z"],["dc.date.available","2017-09-07T11:49:26Z"],["dc.date.issued","2007"],["dc.description.abstract","Dronpa is a novel GFP-like fluorescent protein with exceptional light-controlled switching properties. It may be reversibly switched between a fluorescent on-state and a nonfluorescent off-state by irradiation with light. To elucidate the molecular basis of the switching mechanism, we generated reversibly switchable Dronpa protein crystals. Using these crystals we determined the elusive dark-state structure of Dronpa at 1.95-angstrom resolution. We found that the photoswitching results in a cis-trans isomerization of the chromophore accompanied by complex structural rearrangements of four nearby amino acid residues. Because of this cascade of intramolecular events, the chromophore is exposed to distinct electrostatic surface potentials, which are likely to influence the protonation equilibria at the chromophore. We suggest a comprehensive model for the light-induced switching mechanism, connecting a cascade of structural rearrangements with different protonation states of the chromophore."],["dc.identifier.doi","10.1073/pnas.0700629104"],["dc.identifier.gro","3143453"],["dc.identifier.isi","000248650300011"],["dc.identifier.pmid","17646653"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/969"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0027-8424"],["dc.title","Structural basis for reversible photoswitching in Dronpa"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","32317"],["dc.bibliographiccitation.issue","47"],["dc.bibliographiccitation.journal","Journal of Biological Chemistry"],["dc.bibliographiccitation.lastpage","32327"],["dc.bibliographiccitation.volume","283"],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Luehrmann, Reinhard"],["dc.contributor.author","Wahl, Markus C."],["dc.date.accessioned","2018-11-07T11:08:57Z"],["dc.date.available","2018-11-07T11:08:57Z"],["dc.date.issued","2008"],["dc.description.abstract","The yeast pre-mRNA retention and splicing complex counteracts the escape of unspliced pre-mRNAs from the nucleus and activates splicing of a subset of Mer1p-dependent genes. A homologous complex is present in activated human spliceosomes. In many components of the spliceosome, RNA recognition motifs (RRMs) serve as versatile protein-RNA or protein-protein interaction platforms. Here, we show that in the retention and splicing complex, an atypical RRM of the Snu17p (small nuclear ribonucleoprotein-associated protein 17) subunit acts as a scaffold that organizes the other two constituents, Bud13p (bud site selection 13) and Pml1p (pre-mRNA leakage 1). GST pull-down experiments and size exclusion chromatography revealed that Snu17p constitutes the central platform of the complex, whereas Bud13p and Pml1p do not interact with each other. Fluorimetric structure probing showed the entire Bud13p and the N-terminal third of Pml1p to be natively disordered in isolation. Mutational analysis and tryptophan fluorescence confirmed that a conserved tryptophan-containing motif in the C terminus of Bud13p binds to the core RRM of Snu17p, whereas a different interaction surface encompassing a C-terminal extension of the Snu17p RRM is required to bind an N-terminal peptide of Pml1p. Isothermal titration calorimetry revealed 1: 1 interaction stoichiometries, large negative binding entropies, and dissociation constants in the low nanomolar and micromolar ranges for the Snu17p-Bud13p and the Snu17p-Pml1p interactions, respectively. Our results demonstrate that the noncanonical Snu17p RRM concomitantly binds multiple ligand proteins via short, intrinsically unstructured peptide epitopes and thereby acts as a platform that displays functional modules of the ligands, such as a forkhead-associated domain of Pml1p and a conserved polylysine motif of Bud13p."],["dc.description.sponsorship","Max-Planck-Society"],["dc.identifier.doi","10.1074/jbc.M804977200"],["dc.identifier.isi","000260893700018"],["dc.identifier.pmid","18809678"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/52909"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Soc Biochemistry Molecular Biology Inc"],["dc.relation.issn","0021-9258"],["dc.title","An Unusual RNA Recognition Motif Acts as a Scaffold for Multiple Proteins in the Pre-mRNA Retention and Splicing Complex"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","911"],["dc.bibliographiccitation.issue","10"],["dc.bibliographiccitation.journal","Nature Structural & Molecular Biology"],["dc.bibliographiccitation.lastpage","918"],["dc.bibliographiccitation.volume","21"],["dc.contributor.author","Wysoczanki, Piotr"],["dc.contributor.author","Schneider, Cornelius"],["dc.contributor.author","Xiang, ShengQi"],["dc.contributor.author","Munari, Francesca"],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Luehrmann, Reinhard"],["dc.contributor.author","Becker, Stefan"],["dc.contributor.author","Zweckstetter, Markus"],["dc.date.accessioned","2018-11-07T09:34:34Z"],["dc.date.available","2018-11-07T09:34:34Z"],["dc.date.issued","2014"],["dc.description.abstract","The precursor mRNA (pre-mRNA) retention and splicing (RES) complex is a spliceosomal complex that is present in yeast and humans and is important for RNA splicing and retention of unspliced pre-mRNA. Here, we present the solution NMR structure of the RES core complex from Saccharomyces cerevisiae. Complex formation leads to an intricate folding of three components-Snu17p, Bud13p and Pml1p-that stabilizes the RNA-recognition motif (RRM) fold of Snu17p and increases binding affinity in tertiary interactions between the components by more than 100-fold compared to that in binary interactions. RES interacts with pre-mRNA within the spliceosome, and through the assembly of the RES core complex RNA binding efficiency is increased. The three-dimensional structure of the RES core complex highlights the importance of cooperative folding and binding in the functional organization of the spliceosome."],["dc.identifier.doi","10.1038/nsmb.2889"],["dc.identifier.isi","000342862000011"],["dc.identifier.pmid","25218446"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/32198"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Nature Publishing Group"],["dc.relation.issn","1545-9985"],["dc.relation.issn","1545-9993"],["dc.title","Cooperative structure of the heterotrimeric pre-mRNA retention and splicing complex"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","13070"],["dc.bibliographiccitation.issue","37"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","13074"],["dc.bibliographiccitation.volume","102"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Stiel, André C."],["dc.contributor.author","Gräter, Frauke"],["dc.contributor.author","Schäfer, Lars V."],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:54:19Z"],["dc.date.available","2017-09-07T11:54:19Z"],["dc.date.issued","2005"],["dc.description.abstract","Proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state bear enormous potential in diverse fields, such as data storage, in vivo protein tracking, and subdiffraction resolution light microscopy. However, these proteins could hitherto not live up to their full potential because the molecular switching mechanism is not resolved. Here, we clarify the molecular photoswitching mechanism of asFP595, a green fluorescent protein (GFP)-like protein that can be transferred from a nonfluorescent \"off\" to a fluorescent \"on\" state and back again, by green and blue light, respectively. To this end, we establish reversible photoswitching of fluorescence in whole protein crystals and show that the switching kinetics in the crystal is identical with that in solution. Subsequent x-ray analysis demonstrated that upon the absorption of a green photon, the chromophore isomerizes from a trans (off) to a cis (on) state. Molecular dynamics calculations suggest that isomerization occurs through a bottom hula twist mechanism with concomitant rotation of both bonds of the chromophoric methine ring bridge. This insight into the switching mechanism should facilitate the targeted design of photo-switchable proteins. Reversible photoswitching of the protein chromophore system within intact crystals also constitutes a step toward the use of fluorescent proteins in three-dimensional data recording."],["dc.identifier.doi","10.1073/pnas.0502772102"],["dc.identifier.gro","3143804"],["dc.identifier.isi","000231916300014"],["dc.identifier.pmid","16135569"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1359"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0027-8424"],["dc.title","Structure and mechanism of the reversible photoswitch of a fluorescent protein"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","14603"],["dc.bibliographiccitation.issue","19"],["dc.bibliographiccitation.journal","Journal of biological chemistry"],["dc.bibliographiccitation.lastpage","14609"],["dc.bibliographiccitation.volume","285"],["dc.contributor.author","Brakemann, Tanja"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Groenhof, Gerrit"],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:46:04Z"],["dc.date.available","2017-09-07T11:46:04Z"],["dc.date.issued","2010"],["dc.description.abstract","Reversibly switchable fluorescent proteins can be repeatedly photoswitched between a fluorescent and a nonfluorescent state by irradiation with the light of two different wavelengths. The molecular basis of the switching process remains a controversial topic. Padron0.9 is a reversibly switchable fluorescent protein with \"positive\" switching characteristics, exhibiting excellent spectroscopic properties. Its chromophore is formed by the amino acids Cys-Tyr-Gly. We obtained high resolution x-ray structures of Padron0.9 in both the fluorescent and the nonfluorescent states and used the structural information for molecular dynamics simulations. We found that in Padron0.9 the chromophore undergoes a cis-trans isomerization upon photoswitching. The molecular dynamics simulations clarified the protonation states of the amino acid residues within the chromophore pocket that influence the protonation state of the chromophore. We conclude that a light driven cis-trans isomerization of the chromophore appears to be the fundamental switching mechanism in all photochromic fluorescent proteins known to date. Distinct absorption cross-sections for the switching wavelengths in the fluorescent and the nonfluorescent state are not essential for efficient photochromism in fluorescent proteins, although they may facilitate the switching process."],["dc.identifier.doi","10.1074/jbc.M109.086314"],["dc.identifier.gro","3142927"],["dc.identifier.isi","000277299700057"],["dc.identifier.pmid","20236929"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/385"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Deutsche Forschungsgemeinschaft"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0021-9258"],["dc.title","Molecular Basis of the Light-driven Switching of the Photochromic Fluorescent Protein Padron"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.issue","12"],["dc.bibliographiccitation.journal","MPIbpc news"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Stiel, André C."],["dc.contributor.author","Gräter, Frauke"],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Schäfer, Lars V."],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Grubmüller, Helmut"],["dc.date.accessioned","2018-04-23T08:45:37Z"],["dc.date.available","2018-04-23T08:45:37Z"],["dc.date.issued","2005"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/13255"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.title","Insight into the structure and mechanism of the reversible photoswitch of a fluorescent protein: A multi-departmental research approach"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","35"],["dc.bibliographiccitation.journal","Biochemical Journal"],["dc.bibliographiccitation.lastpage","42"],["dc.bibliographiccitation.volume","402"],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Wahl, Markus C."],["dc.date.accessioned","2017-09-07T11:49:51Z"],["dc.date.available","2017-09-07T11:49:51Z"],["dc.date.issued","2007"],["dc.description.abstract","RSFPs (reversibly switchable fluorescent proteins) may be repeatedly converted between a fluorescent and a non-fluorescent state by irradiation and have attracted widespread interest for many new applications. The RSFP Dronpa may be switched with blue light from a fluorescent state into a non-fluorescent state, and back again with UV light. To obtain insight into the underlying molecular mechanism of this switching, we have determined the crystal structure of the fluorescent equilibrium state of Dronpa. Its bicyclic chromophore is formed spontaneously from the Cys(62)Tyr-(63)Gly(64) tripeptide. In the fluorescent state, it adopts a slightly non-coplanar cis conformation within the interior of a typical GFP (green fluorescent protein) beta-can fold. Dronpa shares some structural features with asFP595, another RSFP whose chromophore has previously been demonstrated to undergo a cis-trans"],["dc.identifier.doi","10.1042/BJ20061401"],["dc.identifier.gro","3143535"],["dc.identifier.isi","000244284400004"],["dc.identifier.pmid","17117927"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1060"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1470-8728"],["dc.relation.issn","0264-6021"],["dc.title","1.8 Å bright-state structure of the reversibly switchable fluorescent protein Dronpa guides the generation of fast switching variants"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.issue","12"],["dc.bibliographiccitation.journal","MPIbpc News"],["dc.bibliographiccitation.lastpage","4"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Wahl, Markus C."],["dc.contributor.author","Stiel, André C."],["dc.contributor.author","Gräter, Frauke"],["dc.contributor.author","Schäfer, Lars V."],["dc.contributor.author","Trowitzsch, Simon"],["dc.contributor.author","Weber, Gert"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Hell, Stefan Walter"],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2018-02-09T07:06:40Z"],["dc.date.available","2018-02-09T07:06:40Z"],["dc.date.issued","2005"],["dc.description.abstract","Schaltbare Proteine, die sich durch Be- strahlung mit sichtbarem Licht reversibel zwischen einem fluoreszierenden ‚Ein’- und einem nicht-fl uoreszierenden ‚Aus’- Zustand hin und herschalten lassen, sind erst seit wenigen Jahren bekannt. Derzeit sind mit den Proteinen asFP595 und Dron- pa erst zwei Vertreter solcher reversibel schaltbaren fl uoreszierenden Proteine (RSFPs) beschrieben worden. Nichts- destotrotz verspricht diese neue Protein- klasse aufgrund ihrer einzigartigen Eigen- schaften bereits jetzt eine Vielzahl interes- santer Anwendungsmöglichkeiten. Das Protein asFP595, dessen Schalt- mechanismus wir in dieser Studie unter- sucht haben, kommt normalerweise in den Tentakelspitzen der Wachsrose Anemonia sulcata vor, einer Seeanemone, die in den lichtdurchfluteten Flachwasser- bereichen des Mittelmeers und des Nord- atlantiks lebt. asFP595 wird durch Be- leuchtung mit grünem Licht von einem nicht-fluoreszierenden ‚Aus’-Zustand in einen fluoreszierenden ‚Ein’-Zustand ver- setzt. Von diesem ‚Ein’-Zustand fällt es spontan in den ‚Aus’-Zustand zurück, kann aber auch durch Beleuchtung mit blauem Licht zurückgeschaltet werden. Dieses lichtgetriebene Schalten ist rever- sibel und viele Male wiederholbar. In einer gemeinsamen Studie dreier Abteilungen haben wir die molekulare Struktur von asFP595 bestimmt und wesentliche Teile des Schaltmechanismus aufgeklärt. Diese Untersuchungen haben gezeigt, dass nach der Absorption eines grünen Photons das asFP595-Chromophor von einer Trans- in eine Cis-Position isomerisiert. Molekular- dynamik-Rechnungen deuten an, dass das Chromophor bei dieser lichtinduzierten Reaktion eine ‚Hula-Twist’-Bewegung macht. Bei dieser Bewegung dreht sich in erster Linie die Methin-Brücke, wel- che die beiden aromatischen Ringe des Chromophors verknüpft. Tatsächlich ändert das Chromophor während des Hula-Twists seine Position lediglich um 3 x 10-10 Meter. Diese winzige Änderung reicht aus, um aus dem nicht-fluoreszierenden ein fluo- reszierendes Protein zu machen. Diese neuen Erkenntnisse über die Struktur und den Schaltmechanismus des Prote- ins sollten es zukünftig ermöglichen, mit Hilfe von gerichteten molekularbiologi- schen Ansätzen das Protein für weitere Anwendungen zu optimieren. Denkbare Einsatzbereiche eines solchen modifizier- ten asFP595 reichen von der höchstauf- lösenden Fluoreszenzmikroskopie bis hin zum Einsatz als Datenspeicher."],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/12069"],["dc.language.iso","en"],["dc.language.iso","de"],["dc.notes.status","final"],["dc.title","Insight into the structure and mechanism of the reversible photoswitch of a fl uorescent protein"],["dc.title.subtitle","A multi-departmental research approach"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]