93311 - Molecular Signalling Transduction

Course Unit Page

SDGs

This teaching activity contributes to the achievement of the Sustainable Development Goals of the UN 2030 Agenda.

Quality education

Academic Year 2021/2022

Learning outcomes

At the end of the course, the student has knowledge of different pathways of intracellular signaling between biological molecules (proteins, nucleic acids, lipids, metabolites) and in-depth knowledge of the main biochemical mechanisms essential for the maintenance and regulation of cellular homeostasis. The student acquires skills for large-scale experimental approaches for modeling and analyzing the complexity of signaling pathways in the context of cellular functions. The student is able to apply the knowledge in practical contexts, including the identification of molecular drug targets and the interpretation of the effect of genetic variations on the functionalities of the signaling pathways. Finally, the student has the skills to maintain and use cell models to perform different biochemical analysis.

Course contents

Part 1- Lectures

Signaling molecular platforms: i) structural and molecular features of 7TM receptors; structural interactions between 7TM receptors and heterotrimeric G proteins; intracellular signaling mechanisms mediated by heterotrimeric G proteins; ii) molecular events during receptor inactivation: structure and function of arrestin proteins; arrestin as an example of molecular switch and signaling platform; iii) molecular organization of adenylate cyclase and phospholipase C as effectors in signaling transduction; functional and structural interactions between 7TM receptors and tyrosine kinase receptors and their transduction mechanisms.

The second intracellular messengers: i) Microdomains of cAMP: PKA and AKAPs proteins; in vivo measurement of the cAMP; ii) Homeostasis of intracellular Ca2+: Ca2+ as a universal and versatile second messenger; the intracellular Ca2+ toolkit; spatio-temporal aspects of Ca2+mediated signaling; aequorin-and GFP-based sensors for the determination of in vivo Ca2+ levels.

Mitochondria and signaling transduction: i) Ultrastructure versus network: cristae, mitochondrial reticulum and their morphological and functional characteristics; mitochondrial dynamics; basic methodologies for the analysis of the structure and function of mitochondria; ii) Mitochondria as a cellular signalling platform: metabolites versus oncometabolites; molecular pathways activated by mitochondrial signals; iii) Signaling between mitochondria and ER: ultrastructural features of the subcellular compartment MAMs and protein networks involved in its maintenance and in its cellular function; role of MAMs in mitochondrial calcium homeostasis (transport systems, structure and function of the Ca2+ uniport); in vivo measurement of mitochondrial Ca2+ using bio-imaging techniques; MAMs as a platform for the biosynthesis of phospholipids between endoplasmic reticulum and mitochondria.

AMPK and mTOR signaling pathways: i) AMPK and mTOR complexes: structure and cellular functions; ii) signaling mediated by the molecular complexes mTORC1 and mTORC2; iii) molecular interaction between AMPK and AKT in signaling triggered by amino acids and growth factors; iv) AMPK and mTOR complexes as a molecular axis in the regulation of autophagy parhway.

Part 2- Experimental laboratory

The experimental laboratory will introduce students to the use of human cell models and to the analysis of the effects of several molecules on i) cell proliferation by using cell viability assay; ii) molecular and biochemical mechanisms determining the expression levels of specific markers (by using SDS/Page and Western blot approaches); iii) tumorigenic potential in vitro by determining the clonogenic ability; iv) analysis and discussion of the obtained data during the practical laboratory.

Readings/Bibliography

Part 1 and Part 2:

Necessary educational supports

Students will be provided with further recommended bibliographical references (reviews and original papers) and the protocol for practical laboratory for a further deepening of the program contents. The educational supports will be provided as PDF files and will be available to the students on the Virtual Learning Environment (https://virtuale.unibo.it).

Advised educational supports

The teacher advises students to consult the following textbooks for clarifications on the basic concepts of cellular and structural biochemistry: i) Cells - Lewin et al; ii) Protein Structure and Function - Petsko and Ringe; iii) Lehninger Principles of Biochemistry - Nelson and Cox.

Teaching methods

Part 1: The teaching method used is based on lectures during which the contents of the program will be illustrated with PowerPoint slides. Attendance to such lessons is not mandatory but it is highly recommended as the contents of the program will be presented and explained by the teacher and will be discussed with the whole class. This teaching method will facilitate the learning of contents and will allow the achievement of the knowledge and skills by the whole class. Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students.

Part 2: During the practical experience, students will be supervised by the teacher and the academic tutor in charge, with a constant dialogue and discussion aimed at clarifying the analytical approaches, the individual experiments performed and the methodologies used. The practical experience in laboratory will be carried out in single-double place at the workbench by using several and different laboratory equipment. Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students. Attendance at the laboratory is mandatory.

Assessment methods

The learning test assessment of Part 1 and Part 2 of MOLECULAR SIGNALLING TRASDUCTION teaching will consist in an interview-test in order to verify and evaluate the student's knowledge of the contents developed and discussed during the lessons and the experimental laboratory. Moreover, the student's ability to integrate and link the different topics with particular attention to the use of scientific terminology and to the proper and accurate exposure will be evaluated. In order to obtain a final grade of 30/30 with honors, the student must show to know in depth all the topics covered during the lessons. Further, the student must explain and integrate the topics with properties of scientific language. To obtain a final grade of 30/30, the student must explain contents covered during the lessons and show the ability to properly integrate them with properties of scientific language. The final grade will be scaled from 30/30 to 18/30 based on the number of questions to which the student is able to answer and on her/his ability to integrate the topics with properties of scientific language. In particular, to obtain the minimum grade of 18/30 the student must show to have basic knowledge of all the contents discussed during the lessons and not be able to integrate them with properties of scientific language. The vote will be considered valid within the calendar year and will contribute to the final mark. The final mark will be determined as the weighted average of the two teaching modules of the MOLECULAR INTERACTION NETWORKS integrated course. Each student will be notified of the final vote via e-mail by Prof. Porcelli, who is responsible of this I.C.

Prof. Porcelli will proceed with the recording of final mark, only once each student has expressly stated by e-mail (annamaria.pporcelli@unibo.it) that he/she accepts the final mark.

Teaching tools

Part 1 and Part 2:

Students are encouraged to communicate to the teacher any requirements by e-mail as soon as possible. This will allow the teacher to evaluate which teaching support tools are most adequate to make the training course accessible to all students. The contents of the lectures will be presented using PowerPoint slides and will be discussed with the whole class through appropriate teaching supports.

Office hours

See the website of Anna Maria Porcelli