Practical Course HP-F8: Components and Mechanisms of Signal Transduction

Type: Practical Course with Student Seminars

Date: 6.-30. April 2020

(no course on 14.-17. April)

Hosts: Ilse Hofmann, Ursula Klingmüller, Tobias Dick and co-workers (responsible organizer contact: i.hofmann@dkfz.de)

Topics:

Cancer cells fail to obey the social constraints that normally maintain tissue organization: they proliferate when they should not, survive where they should not, and invade regions that they should keep out of. Often this is because they have a reduced dependence on signals from other cells for growth, survival, and division. The aim of this course is to gain an insight into problems arising in the field of signal transduction and how they can be addressed.

Content:

Week 1: Cell Adhesion and Signaling (Ilse Hofmann and co-workers)

Over the past decade, several studies on the subcellular distribution of the plaque proteins of adhering junctions have revealed that a number of such proteins are not only constituents of cell-cell contact structures but are also found dispersed in the cytoplasm and nucleus. This dual location suggests that in addition to establishing and maintaining cell adhesive functions these proteins may also play roles in nuclear and ribonucleoprotein processing mechanisms. To identify novel interaction partners and to get an idea on putative signaling cascades involved the following methods can be used. Using different extraction protocols, analytical and preparative immuno-selection experiments followed by mass spec finger printing associated proteins can be identified. Moreover, RNase treatment or RNA-immunprecipitation together with RT-PCR allow the identification of bound RNAs. In addition, the colocalization is investigated by double immunofluorescence microscopy using a confocal laser scanning microscope.

 

Week 2: Systems Biology and Signaling (Ursula Klingmüller, Piotr Zadora, Marcel Schilling, Lorenza D'Alessandro)

Complex intracellular signaling networks mediate signal transmission from cell surface receptors to the nucleus and regulate the activation of transcriptional programs and subsequent cellular responses. Many components of signaling networks have been identified but it remains to be elucidated how information is processed and how cellular decisions are regulated. To address these questions it is important to examine the characteristic dynamic behavior of multiple components of signaling networks simultaneously and combine quantitative data generation with mathematical modeling. Dynamic activation of signaling components by growth factor stimulation at the cell population level will be performed by quantitative immunoblotting and mass spectrometry. In parallel, the impact of growth factors stimulation on cell proliferation will be monitored by measuring DNA synthesis. The generation of a dynamic pathway model and data-based parameter estimation will be demonstrated in a computer practical.

 

Week 3: Oxidative Signaling (Tobias Dick and co-workers)

Reactive oxygen species (ROS) and oxidative stress have long been recognized to be associated with tissue damage and age-related disorders, including malignant, neurodegenerative and cardiovascular disease. More recently, it has also become clear that moderate levels of ROS play essential roles as cellular messengers, causing reversible modifications (thiol oxidation) in specific target proteins. For example, transient oxidative modification of transcription factors contributes to the regulation of inflammatory and stress responses. The fleeting and labile nature of oxidants and intracellular redox states has been a major limitation for research. We will learn about new techniques which allow to identify proteins and pathways regulated by oxidation-reduction processes.

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