Achille Pellicioli
Achille Pellicioli
affiliation: Università di Milano
research area(s): Molecular Biology, Cancer Biology
Course: Biomolecular Sciences
University/Istitution: Università di Milano
Achille Pellicioli had his degree in 1996 in Biological Science with 110/110 summa cum laude at the University of Milano. In 2000 he obtained his PhD in Genetics at
the same University. From 2001 to 2002 he had a postdoctoral fellowship in IFOM-FIRC Institute of Molecular Oncology in the laboratory of Prof. Marco Foiani. In 2002 he obtained a permanent position as a Researcher at the University of Milano working at IFOM- FIRC Institute of Molecular Oncology until February 2006 and from March 2006 at Dipartimento di Scienze Biomolecolari e Biotecnologie, Università di Milano, where he is teaching Molecular Biology.
As documented by several punlications, Achille Pellicioli establised national and intenational collaborabiotions. In particular, in 2002 he visited for collaboration the laboratory of Prof. J. Haber (Brandeis University, Boston), and in 2010 the laboratory of Prof. H. Klein (NYU, New York).
The scientific activity of Achille Pellicioli has been focused on the study of mechanisms of checkpoint signal transduction in response to DNA damage and replication stress, focusing to the checkpoint cellular response to Double Stand DNA Breaks and to the connections between DNA Recombination and Replication. The
laboratory of Achille Pellicioli contributed to the identification and characterization of new checkpoint genes in Saccharomyces cerevisiae and developed a number of genetic and biochemical tools and approches to improve our understanding of the checkpoint mechanisms and their cross-talks with other DNA transaction.
Recently, his lab is focused on the characterization of the mechanisms leading to the inactivation of the DNA damage checkpoint. In particular, the laboratory is working on the functional role of Polo kinases in the regulation of cell division and chromosomes stability in the presence of DSBs.
Achille Pellicioli is the author of several publication on well-recognized peer-rewiewed international journals and in 2010 he became a member of the New York
Academy of Sciences.
Fiorani S, Mimun G, Caleca L, Piccini D, Pellicioli A. (2008). Characterization of the activation domain of the Rad53 checkpoint kinase. Cell Cycle, 7(4), p493-499.
Lazzaro F, Sapountzi V, Granata M, Pellicioli A, Vaze M, Haber JE, Plevani P, Lydall D, Muzi-Falconi M. (2008). Histone methyltransferase Dot1 and Rad9 inhibit single-stranded DNA accumulation at DSBs and uncapped telomeres. EMBO J., 27(10), 1502-12.
Lazzaro F., Giannattasio M., Puddu F., Granata M., Pellicioli A., Plevani P.* and Muzi-Falconi M.* (2009) Checkpoint mechanisms at the intersection between DNA damage and repair. DNA Repair, 8, 1055-1067. *Corresponding authors
Diani, L., Colombelli, C., Tamilselvan Nachimuthu, B., Donnianni, R., Plevani, P., Muzi-Falconi, M. and Pellicioli, A. (2009) Saccharomyces Cdk1 phosphorylates checkpoint kinase Rad53 in metaphase influencing cellular morphogenesis. J. Biol. Chem., 284, 32627-32634.
Donnianni, R., Ferrari, M., Lazzaro, F., Clerici, M., Nachimuthu, B. T., Plevani, P., Muzi-Falconi, M. and Pellicioli, P. (2010) Elevated Levels of the polo kinase Cdc5 override the Mec1/ATR checkpoint in budding yeast by acting at different steps of the signaling pathway. PLoS Genet., 6(1):e1000763.doi:10.1371/journal.pgen.1000763
Pellicioli, A. (2010) Novel insights on Cdc5 and checkpoint adaptation come from meiotic cells. Cell Cycle, 9(8), p1461.
Rossio V, Galati E, Ferrari M, Pellicioli A, Sutani T, Shirahige K, Lucchini G, Piatti S. (2010) The RSC chromatin-remodeling complex influences mitotic exit and adaptation to the spindle assembly checkpoint by controlling the Cdc14 phosphatase. J. Cell Biol., 191(5), p981-97.

Project Title:
Functional roles of Polo kinases in chromosomes stability and cell cycle checkpoints.
Several genetic and epigenetic alterations are required to drive normal cells toward malignant transformation. Cancer genomes are highly rearranged and characterized by complex chromosomal alterations targeting loci containing cancer-relevant genes. Therefore, mechanisms controlling genome stability are relevant to prevent cancerogenesis. In response to DNA damage, all eukaryotic organisms activate a surveillance mechanism, called DNA damage checkpoint, to arrest cell cycle progression and facilitate DNA repair. Several factors are physically recruited to the damaged sites, and specific kinases phosphorylate multiple targets leading to checkpoint activation. Recent studies have shown that elevated levels of the Polo-like kinase 1 (PLK1) act as a tumour-promoting force possibly by down-regulating the checkpoint response. Notably, studies both in yeast and vertebrates have involved Cdc5/PLK1 in turning off the DNA damage checkpoint and promoting the re-start of cell cycle progression after a DNA damage-induced delay. This finding is attractive not only as a potential explanation for the selective pressure allowing outgrowth of malignant cells with acquired defects in the checkpoint network, but also because it suggests potential therapeutic implications. Indeed, a number of agents inhibiting PLK1 have been developed and are currently evaluated as anticancer drugs. However, the selectivity of these PLK1 inhibitors is low, and off-target effects arising through the inhibition of unrelated kinases or inactivation of essential PLK1 functions in normal cells are serious unwanted consequences of such treatments.
To identify genes and factors that may be more specific targets for anticancer therapies, we have set up a biochemical approach in yeast cells to pull down Polo associated factors. Moreover, the outcomes would be important to further characterize the functional roles of Polo kinases in cells responding to DNA damage. We already started to screen protein extracts prepared from yeast cells responding to DSBs and the pulled down proteins would be identified by mass spectrometry (MS/MS). We will also screen protein extracts prepared from cells treated with specific agents (such as MMS, metyl methanesulfonate, and campthotecine) that cause replication stress.
Budding yeast is an ideal model system to screen and study the functional role of novel factors involved in DNA damage and cell cycle checkpoints. Indeed, it is expected that many of the Polo kinase interactors/substrates identified in yeast may have a counterpart in human cells. Therefore, we aim to apply the same biochemical approach to screen for PLK1 interactors/substrates in human cells. We will produce an affinity column with the PBD of PLK1 purified from E. coli cells. Protein extracts from various human cells lines will be prepared after cells treatment with agents that cause DSBs (ionizing radiations, campthotecin, zeocine) and will be analyzed by the affinity column with the PLK1_PBD. The pulled down proteins will be identified by MS/MS.

During the ongoing screening procedure described above, we will start the biochemical and genetic characterization of those proteins identified both in yeast and in human cells. This part of the project is particularly important to evaluate the possibility that the proteins identified may serve as molecular target for cancer therapy. We will take advantage of yeast model system and our experience in yeast molecular biology to start the preliminary characterization of the PLKs interactors.
We will insert a standard epitope in the corresponding gene to study protein stability, localization and putative post-translational modifications in the presence of an irreparable DSB. We will also produce mutations in the corresponding genes to investigate whether they may affect cellular viability in the presence of DNA damaging agents, possibly affecting the Mec1 signaling pathway. Moreover, by taking advantages of specific genetic and molecular approaches, we will analyze checkpoint adaptation and recovery in the obtained mutant strains.
Depending upon the results in yeast, we will then study the orthologs in human cells of the most interesting factors.
We will design specific siRNA to silence the expression of the corresponding genes and we will test cellular viability and checkpoint activation/inactivation in response to treatment with DNA damaging agents. We will also use a specific genetic system, based upon the conditional expression of the endonuclease Sce1 to study checkpoint response, recovery and adaptation to DSBs in human cells.
We will then study post-translational modifications, protein stability and localization in cells responding to DNA damage of the putative PLK1 targets/interactors we identified. Eventually, to visualize and immunoprecipitate the proteins we will evaluate the possibility to use the expression of etherologous proteins carrying the protein fused with standard epitope cassettes.