MOLECULAR BIOLOGY OF EUKARYOTES
Learning outcomes of the course unit
Acquiring knowledge and understanding
The objective is for the students to gain solid and thorough knowledge of the organization of genomes and on the mechanisms and regulation of gene expression in eukaryotic organisms, with emphasis on the emerging roles of noncoding RNA (ncRNA) in gene regulation..
Applying knowledge and understanding
Through guided analysis of key experiments for the molecular understanding of some aspects of eukaryotic gene regulation, the students are expected to strengthen their competence in addressing the experimental study of novel gene expression regulatory pathways and of the involved molecular mechanisms, and the identification and characterization of novel regulatory ncRNAs.
Good knowledge of the structure of nucleic acids and the basic mechanisms of duplications, transcription, repair and recombination of DNA.
Course contents summary
Genomes. Unique sequences, repeated sequences and informative content of eukaryotic genomes; renaturation analysis and identification of genomic components; fraction of genes expressed in a single cell type. Molecular anatomy of a eukaryotic gene: conservation of exons and their structural organization and high intronic variability; evolution of genomes and possible functional significance. Tandem repetition of rRNA genes. Highly repeated sequences and satellite DNA: evolution of satellite DNA mediated by unequal crossing-over events; minisatellites and genetic mapping. Retrovirus, retroposons and interspersed repeated sequences: structure, life cycle and mobilization of retroviruses; retroposons, SINES and LINES sequences and processed pseudogenes. Organellar genomes: circular DNA molecules of mitochondrial and chloroplast genomes. Mapping of genomes. Chromatin. Chromatin, chromosomes and gene activation: the problem of genomic compaction; the nucleosome as the fundamental subunit of chromatin; organization and assembly of octameric histones; phasing nucleosomes, hypersensitive sites; higher order structure of chromatin; centromeres, telomeres and structure of chromosomes. Eukaryotic transcription. Eukaryotic RNA polymerase; eukaryotic promoters; transcription equipment dependent on RNA polymerases I and III; basal transcription apparatus RNA polymerase II dependent; eukaryotic transcription control mechanisms; regulatory "in cis" sequences; regulatory "in transit" factors; DNA binding and transcriptional activation; The various structural protein motifs involved in DNA binding and transcriptional activation. Transcription regulation mechanisms through "enhancers", "silencers" and "insulators"; chromatin structure and its effects on histone code transcription; structural and functional organization of euchromatin and heterochromatin; covalent and non covalent modifiers of chromatin; genomic imprinting. Small RNA. siRNA and RNAi, miRNA, ncRNA and gene regulation. Maturation of RNA. Adjustment of RNA processing mechanisms. Alternative splicing; editing of primary transcripts. Coordination of RNA processing events. Eukaryotic translation: mechanism and regulation.
LODISH H. et al. - BIOLOGIA MOLECOLARE DELLA CELLULA, Italian third ed. Based on the American sixth edition, Zanichelli 2009. Alternatively (original version only) LODISH et al., MOLECULAR CELL BIOLOGY, 7th edition, W.H. Freeman publishers, 2013
WATSON D. et al. - BIOLOGIA MOLECOLARE DEL GENE, sixth edition, Zanichelli 2009
The course consists of lectures on key topics in the program, and in-depth focus on topics of particular relevance and interest, with the use of original scientific articles and the assistance of specialist researchers.
Assessment methods and criteria
Evaluation of the expected achievements will be based on an oral interview/test. The test is devised to evaluate both the molecular-level knowledge of gene expression and regulation mechanisms illustrated during the course, and the ability to apply such a knowledge to addressing and solving experimental problems.