ReTuBi Lecture - Ingrid Grummt
On June 18 of 2018, Ingrid Grummt from DKFZ (Germany) was the invited speaker as an expert visit with the lecture “DNA:RNA triplexes – a liaison between regulatory RNAs and chromatin”.
Summary of the Lecture:
Nuclear long non-coding RNAs (lncRNAs) have been implicated on most, if not all, chromatin-mediated processes. While cataloging the transcriptional output of sequenced genomes has led to new approaches for the discovery of thousands of bona fide lncRNAs, few of these lncRNAs have been ascribed function and very little is known about their mode of action and regulation. We have discovered a novel long non-coding RNA (lncRNA), Khps1, which is transcribed in antisense orientation to the proto-oncogene SPHK1. Khps1 interacts with a homopurine stretch upstream of the transcription start site of SPHK1, forming a DNA:RNA triplex that tethers Khps1 to SPHK1 and guides Khps1-associated p300/CBP to the SPHK1 promoter to local changes of the chromatin structure that activates SPHK1 transcription. Deletion or mutation of the triplex-forming region diminishes enhancer occupancy of E2F1 and p300, attenuates KHPS1-dependent activation of SPHK1 transcription and impairs cell migration and proliferation.
To examine whether lncRNA-mediated triplex formation is a general mechanism of epigenetic regulation, we have established a genome-wide method that allows genome-wide mapping of RNAs that are engaged in DNA:RNA triplex structures. High-throughput sequencing and computational analysis revealed a large set of triplex-forming RNAs originating from non-coding and coding loci harboring super-enhancer and repeat sequences. In a complementary approach, we have identified DNA sequences that are engaged in DNA:RNA triplex structures. Analysis of DNA-associated RNA regions combined with identification of genomic loci that are targeted by triplex-forming RNAs reveals that DNA:RNA triplex formation is a general mechanism of RNA-mediated target-site recognition.
Joint Lab Retreat on Trafficking and Cell Migration in Cancer
From June 6 to 8, 2018 it was held the Joint Lab Retreat on "Trafficking and Cell Migration in Cancer", in Sintra, Portugal. This ReTuBi event had a total of 30 participants from Instituto de Medicina Molecular, Institut Curie and IFOM, the FIRC Institute for Molecular Oncology! Great opportunity to discuss outstanding science!
Cell Migration under confinement Workshop 2018
ReTuBi Lecture - Kristine Schauer
On April 26 of 2018, Kristine Schauer from Institut Curie (France) was the invited speaker as an expert visit with the lecture “Role of actin - and microtubule - dependent motor proteins in Golgi positioning and trafficking”.
Summary of the Lecture:
Many cellular disorders are accompanied by changes in the morphology and positioning of intracellular organelles, yet the functional consequences of organelle alterations are often not clear. The Golgi apparatus is a large and distinctive organelle in mammalian cells constituting the biggest hub of intracellular trafficking at the interface between anterograde and retrograde routes. Although the Golgi apparatus was one of the first organelles to be discovered and observed in detail, the function of its compact structure remains enigmatic. We study cells on bioengineered surfaces of micro-fabricated patterns that provide adhesive cues for defined cell spreading. This approach reduces cell-to-cell variation and mimics space restriction which cells encounter in tissues. We combine this controlled cell culture condition with precise quantification of organelle positioning using a density-based imaging approach. We have previously shown that the Golgi apparatus has a characteristic, well-defined, stable and reproducible organization in micropatterned cells. Because morphology and spatial positioning of organelles are regulated by the cytoskeleton and molecular motor proteins, we are interested in the role of motors at the Golgi. We have used our density-based approach on micropatterns to screen for actin-dependent myosins and microtubule-based kinesins that alter the morphology of the Golgi apparatus. We have identified Myo1C and Kif5B as critical players and have further characterized their role in Golgi positioning, trafficking and cell function. Our analysis will be helpful to understanding the molecular mechanisms underlying intracellular changes associated with cellular pathogenesis such as cancer.
Staff exchange between iMM-Curie (2018/04)
From April 10 to 17 of 2018 Catarina Fonseca from Cláudio Franco (iMM) made a “ReTuBi Staff Exchange” to Franck Perez (Institute Curie) to increase the experience of Catarina Fonseca PhD project which aims to perform a high throughput screening to silence different proteins and observe how is the Golgi position under different conditions using live imaging. To do that, it is necessary to use not only live-imaging techniques as well as use a high throughput analysis that will help to obtain the results. Since FrancK Perez Lab works in Golgi dynamics and already use and establish the high content screening, Catarina Fonseca was able to to get knowledge in this specific technique.
From April 10 to 12 of 2018 Maria Cejudo and Isabela Fortunato from Cláudio Franco (iMM) together with Cláudio Franco made a “ReTuBi Staff Exchange” to Danijela Vignjevic Lab (Institute Curie) to discuss ongoing collaborative projects on gut microvasculature and NM-MIIA function in endothelial cells.
From April 10 to 19 of 2018 João Martins from Edgar Gomes Lab (iMM) made a “ReTuBi Staff Exchange” to Danijela Vignjevic Lab (Institute Curie). The main goal of this staff exchange was to share knowledge about techniques that are required for the development of the projects in each lab. With this, there also the added bonus of being able to interact with many international researchers, improving João Martins communication skills and networking. Danijela Vignjevic’s laboratory has extensive expertise in extracellular matrix alterations by matrix proteinases and co-cultures, which is something that João Martins need to optimize for his PhD project.
From April 10 to 19 of 2018 Raquel Duarte from Luís Costa Lab (iMM) made a “ReTuBi Staff Exchange” to Danijela Vignjevic Lab (Institute Curie). The research project of Raquel Duarte is focus in colorectal cancer. Danijela Vignjevic laboratory at Institut Curie focus on cell migration and invasion, using gut as a model. With this staff exchange, Raquel Duarte was able to learn the techniques used in Danijela Vignjevic laboratory, namely the production of 2D and 3D in vitro cell cultures, that could be then implemented in Luís Costa lab to complement their studies. Is also a great opportunity to discuss results, expand team network and to establish possible collaborations.
From April 10 to 19 of 2018 Francisca Vasconcelos from Cláudio Franco (iMM) made a “ReTuBi Staff Exchange” to Franck Perez (Institute Curie). The Franck Perez Lab studies vesicle intracellular trafficking and has previously establish a molecular system for controlled secretion of cargo proteins (RUSH system) and a high-throughput screening for automated analysis. The staff exchange objective was to define the best strategies to implement a high-throughput screening of drugs that control VEGF secretion using the RUSH system. Thus, Francisca Vasconcelos was able to acquire new technical skills that are important for the development of new research project at Cláudio Franco Lab.
Joint Lab Retreat on Cancer Biology
From April 04 to 06 in Douro (Portugal) it was held the Joint Lab Retreat on Cancer Biology between João Barata Lab (iMM) and François Radvanyi (Institute Curie) with a total of 37 participants.