Genetics and Molecular Biology

The Cell Cycle course aims to provide updated molecular knowledge to address these issues. To this purpose the programme will include two modules: 1. The molecular bases of the cell cycle and its checkpoints 2. The mechanisms of cell division and their importance in the origin and therapy of cancer. MODULE 1. The molecular bases of the cell cycle and its checkpoints The cell cycle engine: concepts and pathways The cell cycle at the heart of vital processes. The cell cycle engine and levels of control. Cyclins, function and conservation: oscillatory molecules and checkpoints. Overview of different regulatory mechanisms: transcriptional, post-transcriptional and proteolysis. Cell cycle entry vs. quiescence Proliferation is often antagonistic to differentiation; G0 and G1 phase are molecularly distinct phases; the restriction point; D-type cyclins; the early part of the cell cycle is largely under transcriptional control; the retinoblastoma protein family; E2F transcription factors; cyclin-dependent kinase inhibitors. Cell cycle in space and time Cell cycle varies from organism to organism and from cell to cell. Cell cycle modulation according to the needs of the cell; changes in cell cycle during Drosophila early development; regeneration and development; functional relevance of polyploidy in health and disease. Senescence checkpoint – when cells get old Types of senescence arrest; activators of senescence; examples of cellular senescence; molecular mechanisms of irreversible arrest; senescence-associated phenotypes including chromatin changes and secretory pathway activation; changes in tissue microenvironment; organismal level; TEAM EXERCISE: Antagonistic pleiotropy of human aging – explore and explain. DNA replication, DNA damage and developmental syndromes - Identification of DNA replication origins; epigenetic control of origin recognition complexes; cell fusion experiments; pre-replication complexes and their activators; the once and only once rule for DNA origin unwinding; many components interplay in initiation of DNA replication; Licensing errors, overeplication and cancer; progression through S phase; cohesins; cohesinopathies; condensins; DNA licensing is coordinated with centrosome licensing. TEAM EXERCISE: Replicating DNA is intrinsically fragile and several types of damage may occur – Present replicative errors and outcomes in details. CRISPR and Repetitive DNA Engineering Special lecture and research overview: Evelyne Tassone (Giunta Lab) MODULE 2. The mechanisms of cell division and their importance in the origin and therapy of cancer Chromatin, nuclear architecture and cell fate Chromatin organization, histone modifications, nuclear architecture, lamin, cohesin complex Preparing chromosome segregation: Antephase, nuclear envelope breakdown and chromosome condensation. Positive feedback loops in Cdk1 activation. Cytoplasmic and nuclear substrates. Restaging of mitosis according to cell cycle activities. Chromosome condensation: structure and role of condensin and cohesion complexes. Sister chromatid resolution and chromosome individualization. Nuclear envelope brake down and nuclear pore disassembly Preparing chromosome segregation: Assembly of the mitotic apparatus and spindle forces. Microtubule structure and their dynamic instability. Minus end and plus end motor proteins. Kinesins and Dynein in mitotic spindle formation. The different pathways for mitotic spindle assembly. Kinetochores and the spindle assembly checkpoint. Point and regional centromeres. Epigenetic determination of centromere function. Constitutive kinetochore proteins and kinetochore assembly at mitosis. Microtubule-kinetochore interactions: from lateral to end on interactions. Microtubule-kinetochore attachments. Genetic, cell biology and biochemical approaches in the discovery of the spindle assembly checkpoint. The role of APC/C in promoting anaphase and its regulation by the SAC. MCC complex formation. Kinetochore-microtubule interactions, error correction and chromosome segregation. Erroneous kinetochore microtubule interactions and their role in SAC. Discovery and consequences of merotelic attachments. The CPC complex: structure, localization and substrates. Aurora B- dependent error correction and the role of tension. Metaphase oscillations and anaphase chromosome movement. Mitotic exit: multiple mechanisms at work. The anaphase promoting complex: structure, multistep activation, early and late substrates. The case of cyclin A. Regulated proteolysis from anaphase to G1. Replication licensing. Counteracting kinase and phosphatase activities at mitotic exit. PP1 and PP2A in mitotic exit. Reformation of the nuclear envelope and of nuclear pores. Reversal of chromosome individualization Mitosis as a therapeutic target in cancer. Microtubule inhibitors in cancer therapy-The complexity of cell fate after microtubule inhibitors. Mitotic slippage and the competing networks model. Apoptotic pathways in mitotically arrested cells and the role of Myc. Other mitotic targets. Inducing high levels of chromosome instability as a therapeutic strategy. Kinetochore-Microtubule attachment stability and cancer therapy. TEAM EXERCISE: Discuss the origins of chemotherapeutic resistance. Mitotic error, aneuploidy, CIN and cancer. The different sources of mitotic errors: checkpoint and cohesion defects, merotelic attachments, pathways to centrosome amplification and tetraploidy. Short term effects of chromosome aneuploidy. Chromotrypsis. P53 activation after chromosome missegregation. Aneuploidy and chromosome instability and their presence in cancer: chromosome and genomic studies. Experimental evidence of the role of chromosome instability in cancer. Human models of chromosome instability and cancer predisposition. Mouse models of mutations in mitotic proteins. Genomic and proteomic changes in aneuploid cells. Aneuploidy associated stresses. TEAM EXERCISE: Draw the BFB cycle in details. DNA integrity checkpoints: local and global response, and their signaling to the mitotic entry machinery. Types of DNA damage; many pathways for DNA repair; the formal organization of the DNA damage response; common players in the DNA replication and DNA repair checkpoints; DNA-defective mutations and checkpoint-defective mutations: Ataxia Telangectasia (and yeast RAD9) are repair-proficient but checkpoint defective; ATM/ATR kinases and their downstream effectors; DNA replication and DNA repair checkpoints: local and global responses; checkpoint systems converge on cyclin B1 regulation. TEAM EXERCISE: Catalog types of DNA damage with associated responses and repair pathways.

Bachelor's Degree in Biological Sciences.

In person classes

Oral examination with presentation of a paper to be agreed upon with professors.

in-presence lectures