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Vögele, Jennifer ; Ferner, Jan ; Altınçekiç, Nadide ; Bains, Jasleen Kaur ; Ceylan, Betül ; Fürtig, Boris ; Grün, J. Tassilo ; Hengesbach, Martin ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; Knežić, Božana ; Löhr, Frank ; Peter, Stephen A. ; Pyper, Dennis Joshua ; Qureshi, Nusrat ; Richter, Christian ; Schlundt, Andreas ; Schwalbe, Harald ; Stirnal, Elke ; Sudakov, Alexey ; Wacker, Anna ; Weigand, Julia ; Wirmer-Bartoschek, Julia ; Wöhnert, Jens ; Duchardt-Ferner, Elke
The SARS-CoV-2 virus is the cause of the respiratory disease COVID-19. As of today, therapeutic interventions in severe COVID-19 cases are still not available as no effective therapeutics have been developed so far. Despite the ongoing development of a number of effective vaccines, therapeutics to fight the disease once it has been contracted will still be required. Promising targets for the development of antiviral agents against SARS-CoV-2 can be found in the viral RNA genome. The 5′- and 3′-genomic ends of the 30 kb SCoV-2 genome are highly conserved among Betacoronaviruses and contain structured RNA elements involved in the translation and replication of the viral genome. The 40 nucleotides (nt) long highly conserved stem-loop 4 (5_SL4) is located within the 5′-untranslated region (5′-UTR) important for viral replication. 5_SL4 features an extended stem structure disrupted by several pyrimidine mismatches and is capped by a pentaloop. Here, we report extensive 1H, 13C, 15N and 31P resonance assignments of 5_SL4 as the basis for in-depth structural and ligand screening studies by solution NMR spectroscopy.
Sreeramulu, Sridhar ; Richter, Christian ; Berg, Hannes ; Wirtz Martin, Maria Alexandra ; Ceylan, Betül ; Matzel, Tobias ; Adam, Jennifer ; Altınçekiç, Nadide ; Azzaoui, Kamal ; Bains, Jasleen Kaur ; Blommers, Marcel Jules José ; Ferner, Jan ; Fürtig, Boris ; Göbel, Michael ; Grün, J. Tassilo ; Hengesbach, Martin ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; Knežić, Božana ; Martins, Jason N. ; Mertinkus, Klara Rebecca ; Niesteruk, Anna ; Peter, Stephen A. ; Pyper, Dennis Joshua ; Qureshi, Nusrat ; Scheffer, Ute ; Schlundt, Andreas ; Schnieders, Robbin ; Stirnal, Elke ; Sudakov, Alexey ; Tröster, Alix Friederike ; Vögele, Jennifer ; Wacker, Anna ; Weigand, Julia ; Wirmer-Bartoschek, Julia ; Wöhnert, Jens ; Schwalbe, Harald
SARS-CoV-2 contains a positive single-stranded RNA genome of approximately 30 000 nucleotides. Within this genome, 15 RNA elements were identified as conserved between SARS-CoV and SARS-CoV-2. By nuclear magnetic resonance (NMR) spectroscopy, we previously determined that these elements fold independently, in line with data from in vivo and ex-vivo structural probing experiments. These elements contain non-base-paired regions that potentially harbor ligand-binding pockets. Here, we performed an NMR-based screening of a poised fragment library of 768 compounds for binding to these RNAs, employing three different 1H-based 1D NMR binding assays. The screening identified common as well as RNA-element specific hits. The results allow selection of the most promising of the 15 RNA elements as putative drug targets. Based on the identified hits, we derive key functional units and groups in ligands for effective targeting of the RNA of SARS-CoV-2.
Wacker, Anna ; Weigand, Julia ; Akabayov, Sabine R. ; Altınçekiç, Nadide ; Bains, Jasleen Kaur ; Banijamali, Elnaz ; Binas, Oliver ; Castillo-Martinez, Jesus ; Çetiner, Erhan Can ; Ceylan, Betül ; Chiu, Liang-Yuan ; Davila-Calderon, Jesse ; Dhamotharan, Karthikeyan ; Duchardt-Ferner, Elke ; Ferner, Jan ; Frydman, Lucio ; Fürtig, Boris ; Gallego, José ; Grün, J. Tassilo ; Hacker, Carolin ; Haddad, Christina ; Hähnke, Martin Jens ; Hengesbach, Martin ; Hiller, Fabian ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; de Jesus, Vanessa ; Jonker, Henry ; Keller, Heiko ; Knežić, Božana ; Landgraf, Tom ; Löhr, Frank ; Luo, Le ; Mertinkus, Klara Rebecca ; Muhs, Christina ; Novakovic, Mihajlo ; Oxenfarth, Andreas ; Palomino-Schätzlein, Martina ; Petzold, Katja ; Peter, Stephen ; Pyper, Dennis Joshua ; Qureshi, Nusrat ; Riad, Magdalena ; Richter, Christian ; Saxena, Krishna ; Schamber, Tatjana ; Scherf, Tali ; Schlagnitweit, Judith ; Schlundt, Andreas ; Schnieders, Robbin ; Schwalbe, Harald ; Simba-Lahuasi, Alvaro ; Sreeramulu, Sridhar ; Stirnal, Elke ; Sudakov, Alexey ; Tants, Jan-Niklas ; Tolbert, Blanton S. ; Vögele, Jennifer ; Weiß, Lena ; Wirmer-Bartoschek, Julia ; Wirtz Martin, Maria Alexandra ; Wöhnert, Jens ; Zetzsche, Heidi
The current pandemic situation caused by the Betacoronavirus SARS-CoV-2 (SCoV2) highlights the need for coordinated research to combat COVID-19. A particularly important aspect is the development of medication. In addition to viral proteins, structured RNA elements represent a potent alternative as drug targets. The search for drugs that target RNA requires their high-resolution structural characterization. Using nuclear magnetic resonance (NMR) spectroscopy, a worldwide consortium of NMR researchers aims to characterize potential RNA drug targets of SCoV2. Here, we report the characterization of 15 conserved RNA elements located at the 5′ end, the ribosomal frameshift segment and the 3′-untranslated region (3′-UTR) of the SCoV2 genome, their large-scale production and NMR-based secondary structure determination. The NMR data are corroborated with secondary structure probing by DMS footprinting experiments. The close agreement of NMR secondary structure determination of isolated RNA elements with DMS footprinting and NMR performed on larger RNA regions shows that the secondary structure elements fold independently. The NMR data reported here provide the basis for NMR investigations of RNA function, RNA interactions with viral and host proteins and screening campaigns to identify potential RNA binders for pharmaceutical intervention.
1H, 13C and 15N chemical shift assignment of the stem-loops 5b + c from the 5′-UTR of SARS-CoV-2
(2022)
Mertinkus, Klara R. ; Grün, J. Tassilo ; Altınçekiç, Nadide ; Bains, Jasleen Kaur ; Ceylan, Betül ; Ferner, Jan ; Frydman, Lucio ; Fürtig, Boris ; Hengesbach, Martin ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; Kim, Jihyun ; Knežić, Božana ; Novakovic, Mihajlo ; Oxenfarth, Andreas ; Peter, Stephen ; Qureshi, Nusrat ; Richter, Christian ; Scherf, Tali ; Schlundt, Andreas ; Schnieders, Robbin ; Schwalbe, Harald ; Stirnal, Elke ; Sudakov, Alexey ; Vögele, Jennifer ; Wacker, Anna ; Weigand, Julia ; Wirmer-Bartoschek, Julia ; Wirtz Martin, Maria A. ; Wöhnert, Jens
The ongoing pandemic of the respiratory disease COVID-19 is caused by the SARS-CoV-2 (SCoV2) virus. SCoV2 is a member of the Betacoronavirus genus. The 30 kb positive sense, single stranded RNA genome of SCoV2 features 5′- and 3′-genomic ends that are highly conserved among Betacoronaviruses. These genomic ends contain structured cis-acting RNA elements, which are involved in the regulation of viral replication and translation. Structural information about these potential antiviral drug targets supports the development of novel classes of therapeutics against COVID-19. The highly conserved branched stem-loop 5 (SL5) found within the 5′-untranslated region (5′-UTR) consists of a basal stem and three stem-loops, namely SL5a, SL5b and SL5c. Both, SL5a and SL5b feature a 5′-UUUCGU-3′ hexaloop that is also found among Alphacoronaviruses. Here, we report the extensive 1H, 13C and 15N resonance assignment of the 37 nucleotides (nts) long sequence spanning SL5b and SL5c (SL5b + c), as basis for further in-depth structural studies by solution NMR spectroscopy.
Schnieders, Robbin ; Peter, Stephen ; Banijamali, Elnaz ; Riad, Magdalena ; Altınçekiç, Nadide ; Bains, Jasleen Kaur ; Ceylan, Betül ; Fürtig, Boris ; Grün, J. Tassilo ; Hengesbach, Martin ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; Knežić, Božana ; Oxenfarth, Andreas ; Petzold, Katja ; Qureshi, Nusrat ; Richter, Christian ; Schlagnitweit, Judith ; Schlundt, Andreas ; Schwalbe, Harald ; Stirnal, Elke ; Sudakov, Alexey ; Vögele, Jennifer ; Wacker, Anna ; Weigand, Julia ; Wirmer-Bartoschek, Julia ; Wöhnert, Jens
The SARS-CoV-2 (SCoV-2) virus is the causative agent of the ongoing COVID-19 pandemic. It contains a positive sense single-stranded RNA genome and belongs to the genus of Betacoronaviruses. The 5′- and 3′-genomic ends of the 30 kb SCoV-2 genome are potential antiviral drug targets. Major parts of these sequences are highly conserved among Betacoronaviruses and contain cis-acting RNA elements that affect RNA translation and replication. The 31 nucleotide (nt) long highly conserved stem-loop 5a (SL5a) is located within the 5′-untranslated region (5′-UTR) important for viral replication. SL5a features a U-rich asymmetric bulge and is capped with a 5′-UUUCGU-3′ hexaloop, which is also found in stem-loop 5b (SL5b). We herein report the extensive 1H, 13C and 15N resonance assignment of SL5a as basis for in-depth structural studies by solution NMR spectroscopy.
Wacker, Anna ; Weigand, Julia ; Akabayov, Sabine R. ; Altınçekiç, Nadide ; Bains, Jasleen Kaur ; Banijamali, Elnaz ; Binas, Oliver ; Castillo-Martinez, Jesus ; Çetiner, Erhan Can ; Ceylan, Betül ; Chiu, Liang-Yuan ; Davila-Calderon, Jesse ; Dhamotharan, Karthikeyan ; Duchardt-Ferner, Elke ; Ferner, Jan ; Frydman, Lucio ; Fürtig, Boris ; Gallego, José ; Grün, J. Tassilo ; Hacker, Carolin ; Haddad, Christina ; Hähnke, Martin Jens ; Hengesbach, Martin ; Hiller, Fabian ; Hohmann, Katharina Felicitas ; Hymon, Daniel ; Jesus, Vanessa de ; Jonker, Henry ; Keller, Heiko ; Knežić, Božana ; Landgraf, Tom ; Löhr, Frank ; Luo, Le ; Mertinkus, Klara Rebecca ; Muhs, Christina ; Novakovic, Mihajlo ; Oxenfarth, Andreas ; Palomino-Schätzlein, Martina ; Petzold, Katja ; Peter, Stephen ; Pyper, Dennis Joshua ; Qureshi, Nusrat ; Riad, Magdalena ; Richter, Christian ; Saxena, Krishna ; Schamber, Tatjana ; Scherf, Tali ; Schlagnitweit, Judith ; Schlundt, Andreas ; Schnieders, Robbin ; Schwalbe, Harald ; Simba-Lahuasi, Alvaro ; Sreeramulu, Sridhar ; Stirnal, Elke ; Sudakov, Alexey ; Tants, Jan-Niklas ; Tolbert, Blanton S ; Vögele, Jennifer ; Weiß, Lena ; Wirmer-Bartoschek, Julia ; Wirtz Martin, Maria Alexandra ; Wöhnert, Jens ; Zetzsche, Heidi