Universitą degli studi di Pavia
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De Nola's curriculum
CURRICULUM
Personal information
Giovanni De Nola CURRICULUM VITAE
Place and date of birth: Sinalunga (SI) 19/June/1988 Address: Via Traversa val di Chiana Ovest n°27
tel: 3922002423
e-mail: giovanni.denola01@universitadipavia.it
Formal Education
2012-today: PhD in genetics and molecular biology
2010-2012: (110/110 cum laude) in “Molecular Biology and Genetics”, University of Pavia - Italy 2007-2010 (105/110) in Molecular Biology (first level) University of Siena- Italy. 2009-2010.
Additional Education
·Partecipation at the IET (Institute of Engineering and Technology) Conference on Synthetic Biology (Cambridge 23-25 March 2008)
·Partecipation at the Fifth World Conference on the Future of Science :“The DNA Revolution” (Venezia 20-22 September 2009).
·Partecipation at the ICGEB theoretical course “RNA structure and function”(Trieste 27 February 1 March 2012).
·Partecipation at the post EURASNET symposium “Regulation of gene expression trough RNA splicing” (Trieste 24-27 March 2012).

Research Related Activities
Jan.2010 - Jul.2010
Position: Specialistic Graduation Candidate (Professor Floriana Rosati) Institute of biochemistry, Siena, Italy
Dec. 2010 – Jul.2012
Position: First Level Graduation Candidate (Supervisor: Dr. Claudia Ghigna)
Institute of Molecular Genetics- National Research Council, Pavia, Italy
The principal interest of my project was to investigate the role of alternative splicing in tumor progression. To this aim we use, as an experimental model, the cancer-specific ΔRon variant of the macrophage-stimulating protein (MSP) receptor.
I was also involved in another project in which we studied how alternative splicing is altered during angiogenesis. I analyzed the expression of Nova2, as well as the alternative splicing profile of its pre-mRNA tergets, in primary endothelial cell cultures (such as human umbilical vein endothelial cells, HUVEC). Moreover, to demonstrate that changes in Nova2 expression levels, as well as modification of alternative splicing of its targets, contribute to angiogenesis process, I have performed(in collaboration with Prof. Elisabetta Dejana; IFOM) Nova2 knockdown studies in endothelial cells using lentiviral vectors carrying shRNAs specific for Nova2.
Further informations
I acquired experience in several molecular and cellular biology techniques, such cloning
techniques, plasmid preparation and purification, classical PCR, PCR-mediated mutagenesis, RT-PCR, electrophoresis on agarose gel, LM gel and acrylamide gel, gel extraction and purification. DNA, RNA and protein quantification. Eukaryotic cell culture. Eukaryotic cell transfection, DNA transfection, RNA extraction from cell pellet and purification.
Additional Skills and Qualifications:
Operating systems: Good knowledge and familiarity with Microsoft Windows XP/Vista and MacOSX (Lion and Snow Leopard)
Microsoft Office package
Adobe Photoshop and Illustrator
Scientific softwares: MacPyMOL, ImageJ, EnzymeX
Bioinformatic tools: NCBI Pubmed, OMIM, GenBank, Entrez Protein, BLAST, EBI ClustalW2, Exon Mine.
Good knowledge of english language
RESEARCH PROJECT
Native structure of the MET receptor tyrosine kinase.
The human genome encodes 59 receptor tyrosines kinases (RTKs) that play fundamental roles in development and tissue homeostasis. Deregulated activity of several RTKs is also a major causative factor in human disease, notably cancer.
The aim of my PhD project is to obtain structural and functional insights of the receptor tyrosine kinase (RTK) MET in its native conformation. MET, like all known RTKs, is a large single-pass membrane protein with a large ectodomain (932 amino acids), a cytoplasmic catalytic domain and flanking sequences involved in regulation of receptor activity, including degradation and recycling.
There are presently numerous crystal structures of the catalytic domain and a large fragment of the ectodomain (amino acids 25-741) but there is no overall structure of the intact receptor. The main purpose of my project is to express native, membrane-bound MET, purify the protein and use cryo-electron microscopy in order to obtain a preliminary, low-resolution map of the native receptor alone and in complex with ligand. Subject to adequate progress in controlling the degree of homogeneity of the native protein MET protein - notably the glycosylation and phosphorylation status - crystallisation experiments will also be undertaken in order to establish the prospects of obtaining 2D and/or 3D crystals that could be used for electron and X-ray diffraction experiments. These studies should offer mechanistic insights into receptor activation and extend our knowledge of the structural basis of cell signalling in higher organisms.
Publications
Personal information
Giovanni De Nola CURRICULUM VITAE
Place and date of birth: Sinalunga (SI) 19/June/1988 Address: Via Traversa val di Chiana Ovest n°27
tel: 3922002423
e-mail: giovanni.denola01@universitadipavia.it
Formal Education
2012-today: PhD in genetics and molecular biology
2010-2012: (110/110 cum laude) in “Molecular Biology and Genetics”, University of Pavia - Italy 2007-2010 (105/110) in Molecular Biology (first level) University of Siena- Italy. 2009-2010.
Additional Education
·Partecipation at the IET (Institute of Engineering and Technology) Conference on Synthetic Biology (Cambridge 23-25 March 2008)
·Partecipation at the Fifth World Conference on the Future of Science :“The DNA Revolution” (Venezia 20-22 September 2009).
·Partecipation at the ICGEB theoretical course “RNA structure and function”(Trieste 27 February 1 March 2012).
·Partecipation at the post EURASNET symposium “Regulation of gene expression trough RNA splicing” (Trieste 24-27 March 2012).

Research Related Activities
Jan.2010 - Jul.2010
Position: Specialistic Graduation Candidate (Professor Floriana Rosati) Institute of biochemistry, Siena, Italy
Dec. 2010 – Jul.2012
Position: First Level Graduation Candidate (Supervisor: Dr. Claudia Ghigna)
Institute of Molecular Genetics- National Research Council, Pavia, Italy
The principal interest of my project was to investigate the role of alternative splicing in tumor progression. To this aim we use, as an experimental model, the cancer-specific ΔRon variant of the macrophage-stimulating protein (MSP) receptor.
I was also involved in another project in which we studied how alternative splicing is altered during angiogenesis. I analyzed the expression of Nova2, as well as the alternative splicing profile of its pre-mRNA tergets, in primary endothelial cell cultures (such as human umbilical vein endothelial cells, HUVEC). Moreover, to demonstrate that changes in Nova2 expression levels, as well as modification of alternative splicing of its targets, contribute to angiogenesis process, I have performed(in collaboration with Prof. Elisabetta Dejana; IFOM) Nova2 knockdown studies in endothelial cells using lentiviral vectors carrying shRNAs specific for Nova2.
Further informations
I acquired experience in several molecular and cellular biology techniques, such cloning
techniques, plasmid preparation and purification, classical PCR, PCR-mediated mutagenesis, RT-PCR, electrophoresis on agarose gel, LM gel and acrylamide gel, gel extraction and purification. DNA, RNA and protein quantification. Eukaryotic cell culture. Eukaryotic cell transfection, DNA transfection, RNA extraction from cell pellet and purification.
Additional Skills and Qualifications:
Operating systems: Good knowledge and familiarity with Microsoft Windows XP/Vista and MacOSX (Lion and Snow Leopard)
Microsoft Office package
Adobe Photoshop and Illustrator
Scientific softwares: MacPyMOL, ImageJ, EnzymeX
Bioinformatic tools: NCBI Pubmed, OMIM, GenBank, Entrez Protein, BLAST, EBI ClustalW2, Exon Mine.
Good knowledge of english language
RESEARCH PROJECT
Native structure of the MET receptor tyrosine kinase.
The human genome encodes 59 receptor tyrosines kinases (RTKs) that play fundamental roles in development and tissue homeostasis. Deregulated activity of several RTKs is also a major causative factor in human disease, notably cancer.
The aim of my PhD project is to obtain structural and functional insights of the receptor tyrosine kinase (RTK) MET in its native conformation. MET, like all known RTKs, is a large single-pass membrane protein with a large ectodomain (932 amino acids), a cytoplasmic catalytic domain and flanking sequences involved in regulation of receptor activity, including degradation and recycling.
There are presently numerous crystal structures of the catalytic domain and a large fragment of the ectodomain (amino acids 25-741) but there is no overall structure of the intact receptor. The main purpose of my project is to express native, membrane-bound MET, purify the protein and use cryo-electron microscopy in order to obtain a preliminary, low-resolution map of the native receptor alone and in complex with ligand. Subject to adequate progress in controlling the degree of homogeneity of the native protein MET protein - notably the glycosylation and phosphorylation status - crystallisation experiments will also be undertaken in order to establish the prospects of obtaining 2D and/or 3D crystals that could be used for electron and X-ray diffraction experiments. These studies should offer mechanistic insights into receptor activation and extend our knowledge of the structural basis of cell signalling in higher organisms.
Publications
