The EMBO Meeting

The EMBO Meeting 2012

 

 

 

 

Monday, 18 Nov 2013

RNA life – from birth to death

 

Sunday, 23 September 09:00-10:30, Apollon Auditorium


David Tollervey

Wellcome Trust Centre for Cell Biology, University of Edinburgh

Lighting up RNA interactions in living cells

In vivo analyses of RNA processing are impeded by two major problems: 1) The kinetics of the process must generally be inferred from steady-state levels of intermediates, so rapid processing events are essentially invisible, particularly if they occur cotranscriptionally. 2) The structures of RNA-protein complexes, within which almost all RNA processing and RNP assembly steps occur, are generally very poorly characterized. These limitations are particularly acute for low abundance members of complex populations, such as pre-mRNAs or ncRNAs. The Tollervey group has therefore been prioritizing techniques to overcome these limitations. Fast kinetic labeling supported by mathematical modeling to analyses rapid processing. UV crosslinking (CRAC) to determine sites of RNA-protein interaction. Crosslinking, ligation and sequencing of hybrids (CLASH) to experimentally identify RNA-RNA interactions. These techniques were developed to understand fundamental RNA metabolism in budding yeast – but are being applied in systems from pathogenic E.coli to virus-infected human cells.

Biography

David Tollervey is Director of the Wellcome Trust Centre for Cell Biology and Professor of RNA Biology at the University of Edinburgh. His group is studying the mechanism and regulation of RNA processing and RNA quality control. Most of the work involves molecular genetic approaches in yeast backed up by biochemistry. David Tollervey took his PhD in Genetics at Cambridge in the lab of Herb Arst, and then moved to the University of California, San Francisco, as a postdoctoral fellow in the laboratory of Chris Guthrie. In 1983 he relocated to a permanent post at the Institut Pasteur in Paris, which he left in 1988 to move to the European Molecular Biology Laboratory (EMBL) in Heidelberg as a group leader. He has been at the University of Edinburgh since 1997. He is a Fellow of the Royal Society and member of EMBO.


Elisa Izaurralde

Department of Biochemistry, Max Planck Institute for Developmental Biology, Tübingen

Regulation of mRNA expression by miRNAs and RNA-binding proteins

The focus of our research is the elucidation of the molecular mechanisms that regulate gene expression at the post-transcriptional level. In particular, we study three specific aspects of this regulation: the mechanisms of miRNA-mediated gene silencing, mRNA quality control and degradation, as well as translational regulation by RNA-binding proteins. Our studies on the mechanisms of miRNA-mediated gene silencing has led to the identification of the GW182 proteins, which are essential for silencing miRNA targets in animal cells. GW182 proteins interact directly with the cytoplasmic poly(A)-binding protein (PABPC) and with the PAN2-PAN3 and CCR4-NOT deadenylase complexes. These interactions are required for miRNA-mediated silencing. The CCR4–NOT complex is a master post-transcriptional regulator in eukaryotic cells. In addition to the GW182 proteins, several RNA-binding proteins have been shown to accelerate deadenylation of their targets by recruiting this complex via protein-protein interactions. Furthermore, the complex is required for bulk mRNA degradation through the 5’-to-3’ mRNA decay pathway. These observations indicate that miRNAs and sequence-specific RNA-binding proteins interface with the general mRNA decay machinery, establishing a complex regulatory network that contributes to cell type and organism-specific gene expression patterns.

Biography

Dr. Elisa Izaurralde was born in Montevideo, Uruguay. She earned her PhD in Biochemistry from the University of Geneva in 1989 before working as a postdoctoral fellow and then as a group leader at the EMBL in Heidelberg. She joined the Max Planck Society as a Scientific Director of the Department of Biochemistry at the Max Planck Institute for Developmental Biology in Tübingen, in October 2005. Dr. Izaurralde is distinguished for her contributions to the understanding of the molecular mechanisms underlying post-transcriptional gene regulation. Dr. Elisa Izaurralde was elected member of EMBO in 2000. She is a recipient of the Friedrich Miescher Award of the Swiss Society for Biochemistry, of the Gottfried Wilhelm Leibniz Prize of the German Research Foundation and of the Ernst Jung Prize for Medicine of the Jung Foundation for Science and Research.


Robert H. Singer

Albert Einstein College of Medicine, Yeshiva University

Following Single mRNAs from Birth to Death in Living Cells

Live cell imaging has been instrumental in analyzing the dynamic properties of RNA. New technologies in optical microscopy and fluorescent probe development have been pushing the envelope of our analysis capabilities. We have been dedicated to developing and implementing these technologies to further the understanding of single mRNA dynamics in cells and organisms. We have utilized computational approaches to analyze real-time transcription activities of endogenous genes from yeast to human cells. We have employed a plethora of imaging methods, ranging from confocal and multiphoton microscopy, long-term cell imaging, high-speed real-time widefield microscopy, single molecule tracking, and we have developed super-registration microscopy and fluorescence fluctuation analysis. We have investigated key processes of RNA synthesis: initiation, elongation, termination, as well as nuclear pore export, cytoplasmic trafficking, localization and decay. Mathematical modeling allowed us to extract quantitative kinetic parameters that precisely describe these processes in living cells.

Supported by NIH GMS Grants to RH Singer

Biography

Dr. Robert H. Singer received his undergraduate degree in physical chemistry from Oberlin College and his Ph.D. in developmental biology from Brandeis University. He then did postdoctoral work in molecular biology at MIT and the Weizmann Institute in Israel. His career has been focused on the cell biology of RNA, its isolation, detection, expression and translation. A patented in situ hybridization technique his lab developed for detecting RNA in morphologically preserved cells revealed that messenger RNA can localize in specific cellular compartments. This work has given rise to the field of RNA transport and localization, enhanced by Dr. Singer's and his colleagues' development of imaging technology and RNA reporters. His lab has shown that the dynamics of RNA transcription can also be interrogated by live cell imaging, as well as by multiplexed fluorescent probes. In addition, Dr. Singer's laboratory has been instrumental in developing rapid and sensitive microscopy that can study single molecules of RNA in living cells and in devising methods to track them from their site of synthesis to the sites of their function. This technology has implications for understanding of the role of RNA in disease such as cancer metastasis and mental retardation. He holds 12 patents on his work.

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