Hot Topics in Bionanoscience
Course description: Speakers from all over the world are invited to present pedagogical introductions to their field with an emphasis on basic concepts. Apart from an introductory lecture, the participants of this course will have an additional discussion with the invited speaker. During that extra hour, they will discuss a recent paper and the holy grails of the field.
The first hour is a lecture and open for everyone to attend (in this case this BN seminar on "Assembly and elongation dynamics of the SARS-CoV-2 replication-transcription complex: a single-molecule perspective" at 12:30-13:30). The second hour is reserved as a discussion hour with the lecturer for the registered class of PhD students and postdocs.
Audience: Registered PhD students and postdocs (see registration form below).
Credits: Those participants who attended (pro-actively) two Hot Topics sessions will be awarded 1 Graduate School Credit (GSC). So this session will count for 0.5 GSC.
Preparation: PhD students who have registered for the Hot Topics course need to prepare for the session by reading the articles listed below.
Date: Friday 10 March 2023
Speaker: David Dulin (Vrije Universiteit Amsterdam)
Host: Dimphna Meijer
Required reading: Participants are required to prepare for this session by reading the following papers (also as download-able files below):
- The nucleotide addition cycle of the SARS-CoV-2 polymerase (Cell, 2021);
- Inhibition of SARS-CoV-2 polymerase by nucleotide analogs from a single-molecule perspective (eLife, 2021);
- Quantitative parameters of bacterial RNA polymerase open-complex formation, stabilization and disruption on a consensus promotor (Nucleic Acids Research, 2022).
Abstract of the BN seminar: Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) has infected hundreds of millions and killed millions worldwide, largely due to the lack of effective therapeutics. To prepare against future deadly variants and coronavirus zoonosis, we must develop new and specific antiviral drugs.
SARS-CoV-2 expresses 16 non-structural proteins (nsp1-16) encoded in the ~30 kb long viral genome. Most nsp’s assemble into a replication-transcription complex (RTC). The RTC synthesizes all the viral RNAs in the host cell, and is therefore a key target for antiviral drugs. The RTC includes a core, made of the nsp12-polymerase and of the elongation factors nsp7 and nsp8. To this core RTC associates several other nsp’s, such as the nsp13-helicase. Despite the wealth of structural studies reported in the last three years, the precise role of these viral proteins during the core RTC assembly and elongation, as well as the function of nsp13-helicase during RNA synthesis remain unclear. Furthermore, an elongation competent RTC in association with nsp13-helicase has not yet been reported. The lack of knowledge on the structure-function relationship of the different viral proteins forming the RTC impairs the rational design of novel antivirals.
Here, we applied high-throughput single-molecule magnetic tweezers to monitor the core RTC dynamics from assembly to full primer extension in various compositions of the RTC, i.e. nsp7, nsp8, nsp12-polymerase and nsp13-helicase. From these high-spatiotemporal resolution data, we derived new models describing the assembly of the core RTC, the nucleotide addition cycle of the core RTC, and how nsp13-helicase impacts the RTC elongation dynamics. Together, our results provide a new vision of the role of RTC viral proteins, establishing the foundation towards the assembly of a functional and complete CoV RTC.
Registration
You can register for the course by filling in the form below. Your place at the course will be confirmed via email before the start of the Hot Topics session. In case there are too many registrants, a selection will be made based on first-come-first-served.