Yesterday, on OCT 10, 2018, Juliane has defended her doctoral thesis on „Identification and characterization of crRNA processing endoribonucleases and new insights in adaptation and interference during CRISPR-Cas immunity in the model cyanobacterium Synechocystis sp. PCC 6803“ with the rarely awarded highest possible rating summa cum laude. Big congratulations from all Cyanolabbers to this phantastic outcome!
Cyanolab, the Genetics and Experimental Bioinformatics unit at the University of Freiburg, Germany, is searching for a scientist to participate in the DFG-funded Priority Program SPP 2141 “Much more than Defence: The Multiple Functions and Facets of CRISPR-Cas” within the project “Cross-talk and extensive rewiring of CRISPR-Cas systems with the cellular regulatory machinery in Cyanobacteria”. The intended start date is within the last three months of 2018 or early in 2019, or upon negotiation and the position is limited to a duration of 3 years.
For recent relevant publications of our laboratory, see Nature Microbiology 3, 367–377; Trends in Biotechnology 36, 996-1010; Cell Reports (2018) 22, 3377–3384; PLoS ONE (2013), 8(2), e56470.
The project tasks will include the analysis of RNA-protein interactions that have an impact on the prokaryotic immune system, with a focus on the cyanobacterial model organism Synechocystis. The work plan includes the immunoprecipitation, isolation and characterization of ribonucleoprotein complexes, the analysis of transcriptomic datasets and transcription factor binding motifs, the generation and molecular analysis of cyanobacterial mutant strains and the study of intracellular interactions with invader DNA and bacteriophages.
For this we seek a highly motivated candidate with a background in biochemistry, molecular biology or molecular genetics (M.Sc. or diploma level).
The project aims at a fundamental understanding of the interactions between the prokaryotic CRISPR-Cas system and the host cell’s regulatory mechanisms. The work is carried out within an ongoing international collaboration and as part of DFG-funded consortium that involves regular meetings and timely delivery of results.
Cyanolab is located at the University of Freiburg, located in one of the most beautiful regions of Southern Germany in the heart of Europe. Freiburg is known for its high quality of life at the slopes of the Black Forest, close to the Alps, Switzerland and France. The Faculty of Biology at the University of Freiburg is one of the leading Life Science Departments in the country.
Please send your application by Email to: firstname.lastname@example.org
In your application, please include (1) a CV in tabular form, (2) a brief summary of previous research experience and academic qualification, (3) scanned copies of university degrees and (4) contact details for at least one reference.
Transcription factors of the GntR type possess two domains: a DNA binding domain in the N-terminal part, and a putative ligand-binding domain in the C-terminal section. Upon ligand binding, the protein activity is modified, leading to a change in the regulation of gene expression. Therefore, GntR-type transcriptional regulators are prime candidates to synchronize metabolism and transcription, especially in Cyanobacteria with their versatile pathways for photosynthesis-driven carbon and nitrogen assimilation. With their new paper “The GntR family transcriptional regulator PMM1637 regulates the highly conserved cyanobacterial sRNA Yfr2 in marine picocyanobacteria” the cyanolabbers Joke, Mascha and Claudia provide for the first time insight into the activity of such a transcription factor from the marine cyanobacterium Prochlorococcus MED4.
Shengwei has defended his doctoral thesis on „Genomic and Transcriptomic Analyses of Marine Microorganisms“ at April 09, 2018. Big congratulations and we wish you success and a good start in the new lab at UCLA!
In our paper “The host-encoded RNase E endonuclease as the crRNA maturation enzyme in a CRISPR–Cas subtype III-Bv system” we show that RNase E is the maturation endoribonuclease of a variant CRISPR system. For the details, please see the publication in Nature Microbiology 3, 367–377 (2018). Please see also our press release. Funding for this research came from the German Research Foundation as part of a grant for the FOR1680 research group: “Unravelling the Prokaryotic Immune System CRISPR-Cas” and the Freiburg Institute for Advanced Studies.
In a joint publication with Shoshy Altuvia’s lab at the Hebrew University of Jerusalem we have studied how the small RNA OxyS protects bacterial cells from DNA damage – see our new paper in The EMBO Journal (2018) 37: 413–426 “OxyS small RNA induces cell cycle arrest to allow DNA damage repair”. This work was supported by a grant from GIF.
The topic of epigenetics in Cyanobacteria was tackled in our work “Identification of the DNA methyltransferases establishing the methylome of the cyanobacterium Synechocystis sp. PCC 6803”, which has appeared in DNA Research. Especially interesting is that the lack of N4-cytosine methylation affects growth and pigmentation of this unicellular cyanobacterium. This work resulted from our long-standing collaboration with the lab of Martin Hagemann at the University of Rostock and the Max Planck-Genome Center Cologne.
Further studying the model cyanobacterium Synechocystis sp. PCC 6803, we contributed to the identification of a previously unknown regulatory set of genes putatively involved in the process of recovery from iron limitation. This work, which resulted from collaboration with Nir Keren’s group at the Hebrew University in Jerusalem, has appeared in The Plant Journal (2018) 93: 235-245.
Asking the question “How can microbes prioritize their responses to multiple environmental stresses?” we found that the sRNA IsaR1 is involved in integrating the responses to iron limitation and high salinity – see our new paper in Environmental Microbiology: “The iron-stress activated RNA 1 (IsaR1) coordinates osmotic acclimation and iron starvation responses in the cyanobacterium Synechocystis sp. PCC 6803“. IsaR1 is an only 68 nt regulatory sRNA that mainly controls the acclimation of oxygenic photosynthesis to iron starvation, details here. This work resulted from our collaboration with the Plant Physiology lab at the University of Rostock and the “Applied Metabolome Analysis” group at the MPI in Golm. Stephan, who has been leading this project is now at the Centre for Environmental Research. Congratulations!
Marine bacteria and Cyanobacteria were studied in another 3 publications. In “Benefit from decline: the primary transcriptome of Alteromonas macleodii str. Te101 during Trichodesmium demise” we investigated how a marine copiotroph benefits from photosynthesis in the co-occurring Trichodesmium. For the details, see our full paper in ISME J. presenting the results of a great collaboration between 4 labs in Germany, Israel and Spain.
In a another great collaboration, with Kaarina Sivonen’s lab in Helsinki, we have investigated Nodularia spumigena, a nitrogen-fixing cyanobacterium that forms toxic blooms in the Baltic Sea each summer. The results from this work are presented in two papers, which appeared in The ISME Journal and in Frontiers in Microbiology. Of special interest are the findings that these bloom-forming cyanobacteria degrade methylphosphonate and release methane.
This month our paper “Acclimation of oxygenic photosynthesis to iron starvation is controlled by the sRNA IsaR1” has been published in the latest issue of Current Biology. IsaR1 is a regulatory RNA molecule that controls the acclimation of cells and especially of the photosynthetic machinery to limiting concentrations of bio-available iron. Please see also our press release and this comment. This work has been a productive collaboration among laboratories in 5 different countries.
In September and October 2016 Cyanolab contributed to five publications.
Research on CRISPR RNA endonucleases: In our article „Structural constraints and enzymatic promiscuity in the Cas6-dependent generation of crRNAs“ in Nucleic Acids Research we investigated two CRISPR-associated RNA endonucleases of the Cas6 family. Small RNA molecules generated by the activity of Cas6 enzymes are an essential element of all CRISPR-Cas systems in the recognition of invading phage DNA or other nucleic acids. However, many bacteria and archaea possess multiple systems and several Cas6 endonucleases. It is poorly understood what makes these enzymes able to keep their substrates apart from each other. We found that the enzymes Cas6-1 (Slr7014) and Cas6-2a (Slr7068) can, despite their sequence identity of only 16.5 %, recognize the respective non-cognate substrate in vitro but not in vivo. We further found that adjacent spacer sequences can interfere with the formation of the short hairpin element within the substrate RNA that is required for proper recognition. This work was supported by the German Research Foundation (DFG) program FOR1680 “Unravelling the Prokaryotic Immune System” and is a joint publication with the group of Rolf Backofen.
Two articles dealt with the regulation of gene expression in cyanobacteria: In “Awakening of a dormant cyanobacterium from nitrogen chlorosis reveals a genetically determined program” in Current Biology the genetic program is described that governs the awakening of a dormant bacterium from nitrogen chlorosis back to photosynthetically active life. This work was performed in collaboration with partners in Germany, the Czech Republic and Japan, led by Karl Forchhammer at the Institute of Microbiology at the University of Tübingen. The results have wide-ranging implications for the understanding of bacterial persistence and cellular aging. More information can be found also here.
In “CyAbrB2 contributes to the transcriptional regulation of low CO2 acclimation in Synechocystis sp. PCC 6803”, which appeared in Plant and Cell Physiology we studied the function of the transcriptional factor cyAbrB2 in the adjustment of primary carbon and nitrogen metabolism to photosynthetic activity under fluctuating environmental conditions.
Two publications targeted the adaptation of microorganisms to the marine environment: In “Trimethylated homoserine functions as the major compatible solute in the globally significant oceanic cyanobacterium Trichodesmium” in PNAS the chemical compound was identified and the biosynthetic pathway leading to it elucidated that assures robust survival under the osmotically harsh high salinity within the marine environment. Globally, substantial amounts of organic carbon are stored in this compound, homoserine betaine, as it contains three methyl groups and occurs in high intracellular concentrations. Thus, its sudden release during the demise of high-density surface blooms of Trichodesmium will impact the marine microbial community. This work was performed in collaboration with partners in Germany and Israel and was led by Martin Hagemann at the Life Sciences Institute of the University of Rostock. More (in German) can be found also here.
In our article „mdRNAseq analysis of marine microbial communities from the northern Red Sea”, which was published in Scientific Reports, we have employed Differential RNA-Seq to analyze the primary transcriptomes of a complex marine microbiome sample. This technique, not previously used for metatranscriptomics, is powerful for the simultaneous identification of transcriptional start sites (i.e., the suite of active promoters) and the analysis of gene expression at community-wide scale. We identified promoters active in situ for five different pico-planktonic organisms genera belonging to the three domains of life (the SAR11 clade of Alphaproteobacteria, Synechococcus cyanobacteria, Euryarchaeota, Thaumarchaeota, and Micromonas as an example for picoeukaryotic algae), showing the applicability of this approach to highly diverse microbial communities.
In July 2016 four publications have appeared to which Cyanolab has contributed.
New regulatory RNA: In the publication de Porcellinis et al. (2016) we characterize PmgR1, a small RNA that is involved in the regulation of growth under photomixotrophic conditions and glycogen metabolism.
Mechanisms for self-destruction: In Kopfmann et al. (2016) we present an overview on the type II toxin-antitoxin systems in the cyanobacterial model Synechocystis sp. PCC 6803. This organism possesses with 69 type II toxin-antitoxin systems plus seven stand-alone components a startling number of such systems. These systems are dangerous biological entitities: Activation of the toxin may lead to the self-destruction of the cell. Indeed, some of these systems function in the post-segregational killing of cells that have lost the plasmid encoding it. However, for the vast majority of toxin-antitoxin systems the functions have remained unknown, especially when encoded in the chromosome. The comprehensive list of these systems will be helpful in their further functional analysis.
Metatranscriptomic analysis of the near-surface microbiome in the South-West Pacific: Two studies contribute to the deeper understanding of marine microbial communities. The work by Pfreundt et al. (2016a) presents the metatranscriptomic analysis of microbial activities within a large mesocosm and the surrounding lagoon waters in the South-West Pacific, 35 km off the coast near Noumea, New Caledonia, over a time of 23 days. In the study by Spungin et al. (2016), cells of the important nitrogen fixer Trichodesmium were collected and studied during the crash of a bloom-like population. Metatranscriptomic profiling showed the upregulated expression of several putative metacaspase genes, suggesting their involvement in the termination of the dense agglomerations of Trichodesmium. These two papers result from our participation in the VAHINE project.
On July 28 Ulrike (Uli) Pfreundt has defended her doctoral thesis on „The non-coding paradox of Trichodesmium – Lifestyles and strategies in the oligotrophic marine environment“ with the highest possible rating summa cum laude. Big congratulations to this phantastic outcome!
Cyanolab is located at the interface of bioinformatics, experimental RNA biology and microbial systems biology. We participate in the Research Training Group “MeInBio – BioInMe: Exploration of spatio-temporal dynamics of gene regulation using high-throughput and high-resolution methods“, the DFG Priority Program SPP 2002 “Small Proteins in Prokaryotes, an Unexplored World” and the DFG Research Group FOR 1680 “Unravelling the prokaryotic immune system CRISPR-Cas“.
We have a long-standing interest in cyanobacteria and other photosynthetic organisms and their functions in the environment and their biotechnology. Current research activities are centered around the analysis of transcriptomic and epigenomic datasets to characterize regulatory RNAs and epigenetic modifications (see, for instance, our publications Hagemann et al., 2018; Hess et al., 2014; Klähn et al., 2015; Kopf et al., 2014; Kopf and Hess, 2015; Lott et al., 2015; Mitschke et al., 2011; Pfreundt et al., 2015; Rübsam et al., 2018; Voigt et al., 2014; Walworth et al., 2015).
We study CRISPR systems with the aim to understand their functions in antiviral defense and also beyond defense (see, for instance, the publications Behler et al., 2018; Hein et al., 2013; Kieper et al., 2018; Scholz et al., 2013).
On the basis of comparative genomic information, computational and experimental tools have been developed and applied to systematically identify novel regulatory RNAs (antisense and non-coding RNAs) in diverse pro- and eukaryotic organisms and to understand their functions (Barshishat et al., 2018; Georg et al., 2009, 2014 and 2017; Lott et al., 2018; Voß et al., 2009; Wright et al., 2013 and 2014).
Particular research has been applied on marine microorganisms and the marine microbiome (Hou et al., 2018; Kopf et al., 2015; Teikari et al., 2018a, b; Voss et al., 2013). In the characterization of the marine microbiome we have contributed to the VAHINE project by studying the microbial metatranscriptome and community omposition in the South-West Pacific (Pfreundt et al., 2016a,b; Spungin et al., 2016; van Wambeke, 2016).