Precision to the very end: what happens when two replication forks converge during termination?

Lead Research Organisation: Brunel University London
Department Name: Life Sciences

Abstract

DNA encodes the information that provides the basis for all life. For cell division to take place the entire DNA of a cell has to be fully duplicated, ensuring that both the mother and the daughter cell can get one complete copy. In addition, the copy made has to be identical to the original. Any changes to the DNA can potentially be harmful, as they will alter how cells function, or even lead to cell death. On a single cell level, accuracy of the duplication process is normally so high that not a single error is made when the millions of DNA base pairs are copied. This extraordinary high level of accuracy is achieved by a network of different processes that regulate the duplication process and choreograph segregation of the two complete copies into mother and daughter. However, mutations in the DNA can inactivate this network of processes and it is a hallmark of cancer cells that they grow in an uncontrolled way.

We are studying the final stages of the duplication process. Replication of DNA is initiated at specific sites called origins of replication. Two complex machines termed replication forks are recruited to these origins. These replication forks are capable of copying the DNA with high precision. While doing so they move in opposite direction at very high speed until they meet another replication fork coming the opposite way. Our research has revealed that the collision of two fast moving replication forks has the potential to corrupt the DNA and introduce mutations. This cause of mutation is unexpected, because the fusion of two such replication forks is a fundamental process when DNA is copied, and we have identified a number of pathways that can prevent the harmful consequences of fork fusions.

Currently we are using the bacterium Escherichia coli to gain a better insight into the mechanics of such fork fusion events. In E. coli only one such fusion event occurs, as the entire DNA is copied by only two replication forks. In our own cells the duplication process is initiated at hundreds of origins, leading to hundreds of fusion events, making studies much more complex. The relative simplicity of fork fusions in E. coli will allow us to study these events at very high detail with many different tools. For example, we will use recently developed microscopy techniques which allow us to directly visualise and track single replication forks inside living cells to study what happens if forks fuse and to identify how these fusion events can cause corruption of the DNA. Our results will allow us to generate a model of how the duplication process of DNA is normally brought to an orderly completion, which will help us to understand this process in our own cells, and our work will reveal whether it might contribute to the development of cancer and ageing.

Technical Summary

All organisms need to replicate their chromosomes with high fidelity to ensure that the genetic information passed on to the next generation is sufficiently accurate. Chromosome duplication initiates at defined origins, with two replication forks proceeding in opposite directions. DNA replication terminates when a replication fork meets the end of a chromosome or another fork travelling in the opposite direction. We have demonstrated in Escherichia coli that fork fusion events, if not processed correctly, result in surprisingly severe consequences, such as persistent over-replication of the chromosome, increased recombination and chromosome segregation defects. Thus, for the accurate completion of genome duplication the fusion of two converging forks must be carefully controlled, a theme also emerging for the hundreds of fork fusion events in eukaryotic cells.

While we have identified some of the pathologies that arise if fork fusions are not processed correctly, our understanding of the molecular mechanics of fork fusion is still limited. Here we propose to use a combined in vivo and in vitro approach in E. coli to directly analyse the protein dynamics and the DNA intermediates arising at fusing forks. We will investigate how fork fusion intermediates are processed and what happens when this processing goes awry, and we will determine how termination is choreographed in the context of whole chromosome dynamics, segregation and cell division. These analyses will provide a detailed view of replication termination and how the incorrect processing of fork fusions can result in pathologies. Our data will form an important foundation for the understanding of how the hundreds of fork fusions in eukaryotic cells are achieved and how their processing contributes towards maintaining genomic stability. Insight into the factors maintaining genomic integrity is much needed for our understanding of cancer, ageing and many hereditary diseases.

Planned Impact

The described programme will provide fundamental insights into what happens when two complex and fast moving replication forks converge and finally fuse. The fusion of replication forks is a necessity of DNA replication and therefore a fundamental aspect of the cell cycle in all organisms. In addition, the implications of our research address fundamental questions of the evolution of chromosomal architecture and replication speed in pro- and eukaryotes. Our studies will shed light on the mechanisms that have evolved to deal with the intermediates arising as forks fuse to allow duplication of the entire chromosome with a fidelity sufficient to avoid significant corruption of the genomic information.

Our recent research in E. coli has demonstrated that fork fusions can result in pathological consequences such as extensive over-replication of the chromosome, increased recombination and problems with cell cycle progression. It will be important to establish a mechanistic basis of how fork fusions are processed to limit genomic instability, as mistakes made during DNA replication are crucial in the development of genetic disease and other mutation-driven problems such as cancer. Clinicians and scientists with interests in hereditary diseases will therefore benefit from our fundamental studies. The general mechanics of DNA duplication is similar in all living organisms and studies in bacterial model organisms have provided many paradigms for understanding these processes in more complex systems. Currently, little research is carried out on replication fork fusions and the potential impact on genomic stability and our results will significantly contribute towards strengthening the international competitiveness of the research on DNA replication and genomic stability carried out within the UK.

Our studies will also have impact on medical and biotechnological applications. Streptomycetes are an important sources for antibiotics. Their chromosome is normally linear, in contrast to many other bacterial species, but it can circularise. It was noted before that this circularisation results in a significant increase of chromosomal instability and it is very tempting to speculate that this instability is a consequence of aberrantly processed fork fusion intermediates. Our work therefore has the potential to be of relevance for technical applications such as large scale culturing of Streptomycetes for production of antibiotics or other secondary metabolites of biological or chemical relevance. Furthermore, we have identified RecG helicase as one of the key players in defusing potentially harmful fork fusion intermediates. The combined deletion of recG and other genes involved in processing fork fusion intermediates is lethal in E. coli. RecG, while being present in most bacterial species, has no known counterpart in mammalian cells. Thus, the proposed work may be of long-term benefit to pharmaceutical applications aiming to develop new targets for inhibition of pathogenic bacteria. Thus, our studies will have relevance to medicine, agriculture and industry.

The proposed research will combine complex biochemical work, molecular genetics and cell biology studies as well as computer modelling approaches to whole genome replication, resulting in significant cross-disciplinary training of all scientists involved. This will strengthen the scientifically-literate workforce and therefore the international competitiveness of the UK. Understanding how healthy organisms maintain genomic stability and cell division, and what happens when these processes go awry, will have long-term benefits to the health and well-being of the UK population. In addition, all researchers of this project will be well-placed to engage with the public to communicate the links between genomes, mutation and the genetic basis of disease, topics of general interest to the public.

Publications

10 25 50
 
Description Objectives 1 and 2 of the proposal aimed to characterise the DNA intermediates formed and the protein dynamics that occur at two converging replication forks, both in vivo and in vitro.

- We have set out to established a number of different systems which allow tracking of active replisomes in living cells. For some time now we have successfully used fluorescently labelled replisome components to follow progression of replication in living cells. One experimental approach that we explored was the differential labelling of the two different replisomes in living cells. Our data have demonstrated that this is possible in principle. However, it was recently shown by the Reyes-Lamothe lab that replisomes are highly dynamic structures which swap replisomes very frequently (Beattie et al. 2017, eLife 6. pii: e21763). Thus, our original approach of tracking the two replisomse via different colours is not possible, as subunits are exchanged at such a quick rate that colour separation will not be maintained over long periods.

We also proposed we would synchronise fork fusion events by holding one fork at a ter/Tus complex, while the other fork is arrested at a lac repressor/operator array. Upon release of the fork arrested at the lac repressor/operator array forks would fuse within a short period of time at the ter/Tus complex. This experimental procedure also has to be changed. Our data accumulated over the past few months demonstrate that fork fusion events are significantly influenced if ongoing synthesis is arrested either by a protein-DNA complex or by a ter/Tus complex (Midgley-Smith et al., 2018a, NAR 46(15):7701-7715; 2018b, NAR 47(4):1847-1860). To make things even more complicated, most fork fusion events in wild type cells take place between two freely moving forks (Dimude et al., 2016, Genes 7(8). pii: E40).

We have overcome these difficulties by developing robust synchronisation techniques. By using temperature-sensitive alleles of the main replication initiator DnaA we can prevent initiation events from occurring, thereby allowing all ongoing rounds of replication to complete synthesis. A shift to permissive temperature then allows the majority of cells to initiate DNA replication within a relatively short period of time. Ongoing rounds of DNA replication can be tracked and our data indicate that about 90% of all forks fuse within a 15 min window. Thus, we can significantly enrich cells in which forks fuse in a population, which has allowed us to start to investigate specific questions related to the fusion of replisomes in these synchronised cells. We also have started to analyse pathologies that occur if key proteins involved in the processing of fork fusion intermediates are missing.

We have also established an in vitro replication system where we can test various fork fusion scenarios. This has resulted in the a set of results clearly demonstrating that a fork fusion event where one fork is arrested at a ter/Tus complex and a freely moving fork results indeed in very different fork fusion intermediates than a situation where two freely moving forks fuse. These studies are in their final stages and we have started to put together a manuscript, which hopefully will be published in 2020.

- In addition we are making excellent use of cells which contain an additional ectopic replication origin, which, as a consequence, generates ectopic fork fusion sites. We have established strain backgrounds with single ectopic replication origins in a number of different locations and we have even generated a strain in which three active origins, oriC and two ectopic origins, are present. We have now been able to reconstitute an ectopic termination area around the site where forks fuse. This has demonstrated that in the absence of RecG over-replication takes place in different locations of the chromosome if forks are forced to fuse here. These results have been published in the form of two research papers (Midgley-Smith et al., 2018a, NAR 46(15):7701-7715; Dimude et al., 2018a, Genes 9(8). pii: E376).

- Having an experimental system with origins in defined ectopic locations has proven an extremely powerful tool to investigate replication dynamics of forks moving in an orientation opposite to normal. We have started to investigate replication dynamics in these systems in the presence and absence of key accessory helicase, such as Rep and DinG. This work was published in Nucleic Acids Research (Hawkinds et al., 2019, NAR 47(10):5100-5113).

Objective 3 of the proposal aims to address whether fork fusion events are mechanistically linked to other genetic elements required for the late stages of chromosome duplication, such as segregation and cell division, and how fork fusions proceed if they are physically separated from these processes.

- We have observed before that the absence of the RecBCD dsDNA end processing enzymes in E. coli leads to the degradation of DNA in the native termination area, but not in the ectopic termination area in strains with two active replication origins. We have started to investigate the precise nature of the degradation. Our data demonstrate that SbcCD nuclease is the main culprit responsible for the degradation observed, an activity also found at forks stalled at RNA polymerase complexes in the absence of the recombination proteins RecBCD (Dimude et al. 2018b, DNA Repair 70:37-48). The degradation observed is not directly associated with fork fusions, as no degradation is observed in an ectopic fork fusion area. In addition, degradation also occurs if the circular chromosome is stably linearised, which means forks do not fuse but replicate until they meet a chromosome end. However, as the degradation point moves when the fork fusion area is moved there appears to be some link between termination and the degradation observed. However, we have not yet identified the precise initiating event responsible for the degradation. The first experimental part of this project is now completed and the results published (Dimude et al., 2018b, DNA Repair 70:37-48).
Exploitation Route Our work on replication termination over the past 10 years has triggered a whole cascade of papers by a number of different labs and both in prokaryotic and eukaryotic cells. The following papers all report results about termination of DNA replication and directly cite our work: Wendel et al. 2014 PNAS 111(46):16454-9; Wendel et al. 2018 PNAS 115(2):349-354; Kuzminov 2015 PLoS Genetics 12(10):e1006229; Azeroglu et al. 2016 12(2):e1005799; Azeroglu et al. 2017 FEBS Letters 591(8):1101-1113; Sinha et al. 2017 PLoS Genetics 13(10):e1006895; Sinha et al. 2018 PLoS Genetics 14(3):e1007256. In addition, I have recently published a book chapter in the electronic library of Science (eLS, Wiley), a resource aimed at advanced undergraduates and non-specialist readers. The non-specialist nature of this article will help to make the material available to a wider audience.
Sectors Education

Manufacturing

including Industrial Biotechology

Pharmaceuticals and Medical Biotechnology

URL http://www.rudolphlab.com
 
Description Staff training My co-worker employed on the grant was already employed on the previous grant and has been highly trained in all aspects of a modern molecular biology lab as well as training on more specialised equipment such as using an Amnis ImageStream Mark II for high-resolution in-flow microscopy and specialised gel electrophoresis techniques, such as pulsed-field gel electrophoresis and 2D-DNA electrophoresis. Currently I am training her specifically in advanced fluorescence microscopy techniques that will allow her to visualise protein and chromosome dynamics in living cells, one of the important techniques for this specific awards. She was a named postdoctoral researcher on a joint grant proposal with Dr Ed Bolt (Nottingham), which was successful, so I am delighted that she will remain in the lab. This particular grant is heavily reliant on fluorescence microscopy, so she will be able to put her training to excellent use. In addition, we have been working together on developing library preparation procedures from genomic DNA from E. coli for Deep Sequencing in our lab, a skill that is highly relevant and sought after. This is currently still ongoing and also will be relevant for the position she is currently employed on. She has regularly presented her results in lab meetings, both local and with collaborators, and she has just been selected to present her findings at the Annual Conference of the Microbiology Society in Edinburgh as part of the Prokaryotic Genetics and Genomics forum as well as the Young Microbiologists Symposium 2017 in Norwich. These presentation activities will continue to enhance her communication skills, skills that are extremely important for being successful on the current national and international job market. In addition we continue to train undergraduate and postgraduate students at a very high level. Within the running period of this grant four of our undergraduate students have generated data of such high quality that they have been included in the list of authors for two research papers that were published last year (Midgley-Smith et al. 2018, NAR 46(15):7701-7715; Dimude et al. 2018, Genes (Basel) 9(8), pii: E376), a remarkable achievement for students at an undergraduate level. From previous experience we know that these achievements will significantly improve their chances on the job market once they graduate. One of these students has started studies for his PhD at Brunel University, a second student has become a Research Assistant at the Next Generation Sequencing Facility at the Babraham Institute, a position that she will soon leave in order to start studies for her PhD, and two other have graduated with First Class Honours degrees in Summer 2019. In addition, both in March 2017 and 2018 my lab has provided school students with a hands-on experience in molecular cloning and bacterial genetics, ranging from single students visiting the lab to whole classes gaining some practical experience. In addition, I have provided a 4 week work experience period for a school student in autumn 2019, and we are planning to do this on a more regular basis in future years. These events regularly result in students being recruited to study STEM subjects at a University level, resulting not only in them gaining specialist knowledge such as Biomedical Sciences, but also contributing to the wider economy of the UK. Internet As detailed in our Pathways to Impact document, we have developed a lab-specific website to make our research accessible to the interested public: www.rudolphlab.com We spend considerable time developing the content and we have introduced a blog functionality as a major update to provide additional ways of making our research results accessible to specialists and lay people alike. We will present our most recent research results on this platform and we are aiming now to expand into other social and scientific networks such as Researchgate, Publons, Google Scholar and Twitter to attract the widest readership possible.
First Year Of Impact 2018
Sector Education,Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Societal

 
Description Harry Smith Summer Vacation Studentship
Amount ÂŁ1,880 (GBP)
Funding ID GA001445 
Organisation Microbiology Society 
Sector Learned Society
Country United Kingdom
Start 05/2019 
End 07/2019
 
Description Harry Smith Summer Vacation Studentship
Amount ÂŁ1,880 (GBP)
Funding ID VS17/25 
Organisation Microbiology Society 
Sector Learned Society
Country United Kingdom
Start 06/2017 
End 09/2017
 
Description Harry Smith Summer Vacation Studentship
Amount ÂŁ1,880 (GBP)
Funding ID GA000523 
Organisation Microbiology Society 
Sector Learned Society
Country United Kingdom
Start 05/2018 
End 07/2018
 
Title Chromosomal over-replication in cells lacking 3' and 5' exonucleases 
Description Replication profiles generated via Deep Sequencing analysing chromosomal over-replication in E. coli cells lacking the 3' exonucleases ExoI, ExoVII and SbcCD as well as the 5' exonuclease RecJ. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
Impact Raw data are available for other researchers to scrutinise results published and to use the available data for mining in different areas of research, thereby driving forward current research projects. 
URL http://www.ebi.ac.uk/ena/data/view/PRJEB28600
 
Title DNA degradation in the termination area of cells lacking RecBCD 
Description Bacterial chromosome duplication is initiated at a single origin (oriC). Two forks are assembled and proceed in opposite directions with high speed and processivity until they fuse and terminate in a specialised area opposite to oriC. Proceeding forks are often blocked by tightly-bound protein-DNA complexes, topological strain or various DNA lesions. In Escherichia coli the RecBCD protein complex is a key player in the processing of double-stranded DNA (dsDNA) ends. It has important roles in the repair of dsDNA breaks and the restart of forks stalled at sites of replication-transcription conflicts. In addition, ?recB cells show substantial amounts of DNA degradation in the termination area. In this study we show that head-on encounters of replication and transcription at a highly-transcribed rrn operon expose fork structures to degradation by nucleases such as SbcCD. SbcCD is also mostly responsible for the degradation in the termination area of ?recB cells. However, additional processes exacerbate degradation specifically in this location. Replication profiles from ?recB cells in which the chromosome is linearized at two different locations highlight that the location of replication termination can have some impact on the degradation observed. Our data improve our understanding of the role of RecBCD at sites of replication-transcription conflicts as well as the final stages of chromosome duplication. However, they also highlight that current models are insufficient and cannot explain all the molecular details in cells lacking RecBCD. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
Impact No notable impact yet, but the paper was already several times cited in the research literature, highlighting the academic impact. 
URL http://www.ebi.ac.uk/ena/data/view/PRJEB27616
 
Title DNA replication dynamics in Escherichia coli strains with an increasing number of ectopic replication origins 
Description Following on from our oriZ work (Ivanova et al. 2015, Nucleic Acids Res.43(16):7865-77) we have generated strains with an additional ectopic replication origin termed oriX in the replichore opposite to the locatio of oriZ. Replication parameters were analysed in a way similar to our oriZ work, but we also managed to generate a strain background that carried 2 ectopic origins simultaneously, oriC+ oriX oriZ. Replication profiles were generated via Next Generation Sequencing and all Sequencing data will be made publically accessible via the European Nucleotide Archive. 
Type Of Material Database/Collection of data 
Year Produced 2017 
Provided To Others? Yes  
Impact The datasets will allow researchers interested in replication dynamics and chromosome evolution in bacteria to study replication dynamics in cells with a variety of combinations of two, a single ectopic, two ectopic or three replication origins. 
URL http://www.ebi.ac.uk/ena/data/view/PRJEB19883
 
Title DNA replication dynamics in double origin cells lacking Rep helicase 
Description Bacterial genome duplication and transcription require simultaneous access to the same DNA template. Conflicts between the replisome and transcription machinery can lead to interruption of DNA replication and loss of genome stability. Pausing, stalling and backtracking of transcribing RNA polymerases add to this problem and present barriers to replisomes. Accessory helicases promote fork movement through nucleoprotein barriers and exist in viruses, bacteria and eukaryotes. Here we show that stalled E. coli transcription elongation complexes block reconstituted replisomes. This physiologically relevant block can be alleviated by the accessory helicase Rep or UvrD, resulting in the formation of full-length replication products. Accessory helicase action during replication-transcription collisions therefore promotes continued replication without leaving gaps in the DNA. In contrast, DinG does not promote replisome movement through stalled transcription complexes in vitro. However, our data demonstrate that DinG operates indirectly in vivo to reduce conflicts between replication and transcription. These results suggest that Rep and UvrD helicases operate on DNA at the replication fork whereas DinG helicase acts via a different mechanism. 
Type Of Material Database/Collection of data 
Year Produced 2019 
Provided To Others? Yes  
Impact No immediate impact yet, but the related paper is already cited in the scientific literature. 
URL http://www.ebi.ac.uk/ena/data/view/PRJEB20003
 
Title Over-replication in double origin cells lacking RecG helicase 
Description Replication profiles of cells with one native, one ectopic or two replication origins, showing that areas where replication forks fuse are associated with over-replication of the bacterial chromosome 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
Impact The raw data are available for other researchers not only for scrutinising published results, but also for mining the existing data in other context, thereby driving active areas of research forwards. 
URL http://www.ebi.ac.uk/ena/data/view/PRJEB25595
 
Description Bernard Hallet 
Organisation Catholic University of Louvain
Country Belgium 
Sector Academic/University 
PI Contribution Prof. Hallet's group investigates the molecular mechanism of transposon integration into bacterial chromosome and has found links of transposon integration with DNA replication and specifically replication termination. We have provided his lab with a number of strain constructs which will allow him to probe for various aspects of replication and termination. We have organised a Symposium in August 2016 with a variety of groups participating to have an active scientific exchange about links between replication, termination and DNA integration mechanisms.
Collaborator Contribution Exchange of ideas and concepts.
Impact No outputs yet.
Start Year 2016
 
Description Ed Bolt 
Organisation University of Nottingham
Department School of Life Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution My lab is actively working on generating strains in which proteins of the Cas-CRISPR system can be localised in vivo. We are investigating the role of the CRISPR-Cas system in bacteria in general and in E. coli in particular.
Collaborator Contribution Ed Bolt's lab is working on the in vitro reconstitution of the Cas-CRISPR system in Escherichia coli. The collaboration will allow us to combine both in vivo and in vitro approaches to characterise the system in E. coli and other bacterial organisms.
Impact No outputs yet
Start Year 2016
 
Description Mark Leake 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution My lab provides cell biology expertise and research materials.
Collaborator Contribution Mark Leake provides single molecule imaging expertise, access to instrumentation and research materials.
Impact No specific outputs yet.
Start Year 2019
 
Description Michelle Hawkins (York) 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution We are investigating the issues arising from the termination of DNA replication from two separate angles. My lab focuses on the events associated with the fusion of two replication forks in vivo and the consequences that arise if certain protein activities are not present in cells.
Collaborator Contribution Michelle Hawkins is a very good biochemist who was trained in the lab of my former collaborator Peter McGlynn. She has taken over the investigations of the events associated with the fusion of two forks in an in vitro system where DNA replication is reconstituted from purified components.
Impact Research papers: PMID 30869136
Start Year 2018
 
Description Nanna Therapeutics 
Organisation Nanna Therapeutics
Country United Kingdom 
Sector Private 
PI Contribution Exchange of knowledge and ideas
Collaborator Contribution Materials for the generation of an E. coli insertion library, plus relevant protocols
Impact No outputs yet.
Start Year 2019
 
Description Ole Skovgaard 
Organisation Roskilde University
Country Denmark 
Sector Academic/University 
PI Contribution Our lab has contributed a large number of datasets to some data from the Skovgaard lab, leading not only to a publication but also triggering a number of experiments that are currently developed and will result in future publications.
Collaborator Contribution Ole Skovgaard has significantly contributed towards our ongoing research by providing bacterial strains and other materials, but also by contributing towards manuscripts, some of which are already published and some of which are currently defeloped.
Impact Research papers: PMIDs 26160884; 30060465; 32351461
Start Year 2014
 
Description Peter McGlynn 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution Our lab is working on in vivo system of characterising replication fork fusions in living cells.
Collaborator Contribution Peter McGlynn is an excellent biochemist, which complements my own cell biology expertise very well. He is actively working on characterising replication fork fusion in a in vitro system with purified components. Peter McGlynn retired in September 2018, and the collaboration was continued with Dr Michelle Hawkins, a postdoc who was trained in the McGlynn lab.
Impact Research papers: PMIDs 19941825; 20923786; 30869136 Joint BBSRC grant BB/N014995/1 "Precision to the very end: what happens when two replication forks converge during termination?"
Start Year 2007
 
Description Renata Retkute 
Organisation University of Warwick
Country United Kingdom 
Sector Academic/University 
PI Contribution Renata Retkute is an expert for computer modelling of replication dynamics in yeast and bacteria. Her modelling expertise excellently complements my own cell biology knowledge.
Collaborator Contribution Analysis of experimental data, integration of experimental data into computer modelling scenarios, evaluation of modelling results, leading to refind experiments.
Impact Research papers: PMIDs 26160884; 26530381; 30060465
Start Year 2013
 
Description CoolLED interview 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Based on an entry in blog of my internet page (https://rudolphlab.com/blog) the company CoolLED reached out to me to conduct an interview about the research in my lab and how the recent accquisition of a pE-4000 LED illumination unit helps the work in my lab. This was published online and in their monthly newsletter, and links have been posted on Social Media such as Twitter.
Year(s) Of Engagement Activity 2021
URL https://www.coolled.com/interviews/under-the-microscope-10-understanding-dna-replication-dynamics-in...
 
Description DNA integration, replication dynamics and replication termination in Escherichia coli 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I have organised the Symposium "DNA integration, replication dynamics and replication termination in E. coli" at Brunel University London in August 2016, which was attended by 4 research groups from the UK, one research group from Louvain-La-Neuve in Belgium, and a research group from the University of Zagreb in Croatia. All groups have a related but so far distinctly different research focus, but we have recently identified overlapping interests. The Symposium was aimed to develop the overlapping interests further into more formal collaborations between our groups. This approach was very successfull and has resulted in consolidating 2 direct collaborations. The Symposium was much appreciated by all participants and we are discussing whether we can repeat it in 2017 with an increased number of research groups.
Year(s) Of Engagement Activity 2016
 
Description Helicases2021 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Scientific conference with the a focus on helicases and how they contribute to human health and disease. The event was an online conference that was attended by a large number of researchers as well as postgraduate and also undergraduate students.
Year(s) Of Engagement Activity 2021
URL https://premc.org/helicases2021/
 
Description Invited Research Seminar NTU 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other audiences
Results and Impact Invited seminar entitled "Colliding forks - structural parameters of the E. coli chromosome" at the School of Science and Technology, Nottingham Trent University
Year(s) Of Engagement Activity 2016
 
Description Invited oral presentation at the 7th European Conference on Prokaryotic and Fungal Genomics, Göttingen, Germany 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I was invited to present my talk >>Origins left, right and centre: increasing the number of initiation sites in the Escherichia coli chromosome<< in the group section "Synthetic Biology"at the 7th European Conference on Prokaryotic and Fungal Genomics, Göttingen, Germany. Some of the interactions that followed from this talk were with related industries, but also with PIs, PDRAs and PhD students.
Year(s) Of Engagement Activity 2017
URL http://www.prokagenomics.org/
 
Description Invited oral presentation at the Annual Conference of the Microbiology Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact My postdoc, Juachi U. Dimude, was invited to present our data relating to integrating additional replication origins into the Escherichia coli chromosoe at the Prokaryotic genetics and genomics forum as part of the Annual Conference of the Microbiology Society.
Year(s) Of Engagement Activity 2017
URL http://www.microbiologysociety.org/events/annual-conferences/index.cfm/annual-conference-2017
 
Description Invited research seminar at The University of Birmingham 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact As a direct result of my postdocs talk the Annual Meeting of the Society of Microbiology I was invited for a Research Seminar by Prof. Stephen Busby and Prof. David Grainger to present my work at the Institute of Microbiology and Infection, University of Birmingham in October 2017. I had very good interactions with the researchers and PIs at the IMI
Year(s) Of Engagement Activity 2017
 
Description JUD Invited oral presentation at the Young Microbiologists Symposium 2017, John Innes Centre, Norwich, UK 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact My postdoc Dr Juachi U. Dimude was invited to present our work at the Young Microbiologists Symposium 2017, which took place in September 2017 at the John Innes Centre in Norwich. Her talk was entitled: >>A start in many places: Structural insights of Escherichia coli chromosome by introducing an increased number of replication origins<<.
Year(s) Of Engagement Activity 2017
 
Description Research Seminar University of Nottingham Spring 2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other audiences
Results and Impact Invited Research Seminar at the School of Biosciences to introduce the recent findings of my lab.
Year(s) Of Engagement Activity 2018
 
Description Research Seminar University of Warwick Summer 2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Undergraduate students
Results and Impact Research seminar at the School of Medicine, Warwick University, to describe the most recent research findings of our group.
Year(s) Of Engagement Activity 2018
 
Description Research seminar at the University of Nottingham 2019 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other audiences
Results and Impact Research seminar about my research activity in the Faculty of Medicine & Health Sciences, Queen's Medical Centre, University of Nottingham; attended by faculty members and postgraduate students.
Year(s) Of Engagement Activity 2019
 
Description SLM Poster presentation at SoM Annual Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact My PhD student Sarah Midgley-Smith presented her poster on the role of 3' exonucleases in termination of DNA replication in Escherichia coli. The poster session was attended by all participants of the Annual Meeting of the Society of Microbiology and Sarah interacted with a number of researchers and research students.
Year(s) Of Engagement Activity 2017
URL http://www.microbiologysociety.org/events/annual-conferences/index.cfm/annual-conference-2017
 
Description Seminar Okinawa Institute of Science and Technology 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I was invited to present the latest research of my group as a Research Seminar at the Okinawa Institute of Science and Technology, Graduate University, Onna, Okinawa, Japan. I was personally invited by Prof. Simone Pigolotti to give this research seminar. This meeting was attended both in person in via online connections by a large number of researchers from all over Japan and other locations. My talk sparked questions and discussions afterwards.
Year(s) Of Engagement Activity 2022
 
Description Seminar Tokyo Metropolitan 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I was invited to present the latest research of my group as a Research Seminar at the Institute of Medical Science, Tokyo Metropolitan University. I was personally invited by the Director of the Institute of Medical Science, Prof. Hisao Masai, to give this research seminar. This meeting was attended both in person in via online connections by a large number of researchers from all over Japan and other locations. My talk sparked questions and discussions afterwards.
Year(s) Of Engagement Activity 2022