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Keywords
TRANSCRIPTION REGULATION, CHROMATIN REMODELING, HISTONES, PROTEIN COMPLEXES, STRUCTURAL BIOLOGY, LYSINE-SPECIFIC HISTONE DEMETHYLASE 1, C-TERMINAL BINDING PROTEIN, DEVELOPMENT, TUMORIGENESIS

Structural properties and functional activities in a chromatin remodeling nuclear protein complex

Università degli Studi di Milano
Abstract
The chromatin nucleosome core particle is a compact assembly composed of ds-DNA wrapped around the histone octamer. The N-terminal residues of histones protrude from the nucleosome complex, and are target of intensive post-translational modifications (e.g. methylation, acetylation, phosphorylation, and sumoylation). The modifications act in combinatorial and sequence-dependent manners to yield specific downstream events; additionally, they are used as a cellular vocabulary for the regulation of key transcription-based processes. Each cell type is characterized by its own pattern of chromatin marks (the so-called “histone code”). Epigenetics (the study of heritable changes in genome function occurring without change in DNA sequence) aim at explaining how chromatin allows patterns of gene expression to be transferred from one cell to its descendants, and how gene expression changes during differentiation of one cell type into another. These series of events deeply impact on our understanding of human disease, cancer, ageing and stem cells, supporting the growing interest towards “epigenetic therapies” and “epigenetic drugs”. Indeed, regulation of the epigenetic processes has a strong potential for therapy, as shown by the efficacy of an increasing number of inhibitors that target different classes of chromatin-modifying enzymes. In particular, inhibitors of DNA methylation, and of histone deacetylases, are in advanced clinical trials for the treatment of selected forms of >>>

Principal Investigator
Martino Bolognesi Università degli Studi di MILANO
Research Objectives
Main aim of this proposal is to expand our limited knowledge on the structural bases and on the function of selected protein complexes involved in chromatin remodeling and transcriptional regulation. As a result of a successful series of collaborative studies recently carried over by the research units filing this proposal, we address specific questions on the role and properties of two nuclear proteins (CtBP and LSD1) that are involved in histone and chromatin modification. Such studies, on a longer time scale, aim at a deeper understanding of epigenetic processes that are increasingly shown to be related to development, differentiation and tumorigenesis.
Specific activities carried over by the three proposing units (UNIMI-1, UNIPV-2,,UNIMI-3) are detailed in the associated B-forms. The proposal stems form a previous scientific collaboration between UNIPV-2 and UNIMI-3, supported by a PRIN-2004 grant, that brought to the discovery of LSD1 activity. The obvious link to the activities at UNIMI-1, targeting the core component of the CtBP complex, brought to the assembly of this proposal. Previous collaborations between UNIMI-1 and UNIPV-2 are fully documented in the literature (e.g. see www.rcsb.org/pdb).

Aim_1. To elucidate the three dimensional structure of Carboxyl-terminal binding proteins (CtBPs). CtBP are multi-functional proteins implicated in gene regulation, Golgi maintenance and synaptic ribbon formation (Schaeper et al., 1995). When localized >>>

Timescale
24 months
National and international background
Epigenetics is the study of heritable changes in genome function that occur without a change in DNA sequence. This includes the study of how chromatin allows patterns of gene expression to be passed from one cell to its descendants, and how gene expression changes during the differentiation of one cell type into another. The chromatin nucleosome core particle is a compact assembly formed by DNA wrapped around the histone octamer. The flexible N-terminal amino acids of histone tails protrude from the nucleosome and are targets for various post-translational modifications such as methylation, acetylation, phosphorylation, and sumoylation (Grunstein, 1997; Spotswood and Turner, 2002). These modifications act in combinatorial and sequence-dependent manners to yield specific downstream events, and are therefore used as a cellular vocabulary for the regulation of different transcription-based processes (Jenuwein and Allis, 2001). Each cell type is characterized by its own pattern of such chromatin marks, forming the so-called “histone code”.
Epigenetics impacts on our understanding of human disease, cancer, ageing and stem cells; therefore there is a growing interest in the development of so-called “epigenetic therapies” and “epigenetic drugs” (two major reviews on this topic has been published in the first three months of the 2006; Yoo and Jones, 2006; Inche and La Thangue, 2006). A proof-of-concept of the potential of the epigenetic processes in the treatment of >>>