Molecular Mechanisms of Chromosome Dynamics

Fachinetti

Daniele Fachinetti CR1 CNRS Tel:

Figure 1 : Les centromères sont requis pour la séparation des chromosomes lors de la mitose et de la méiose. Immuno-fluorescence sur cellules humaines montrant la localisation de CENP-C et alpha-tubulin.
Figure 1: The centromeres are required for chromosome segregation during mitosis and meiosis. Immunofluorescence on human cells shows the localization of CENP-C and alpha-tubulin during the different stage of mitosis.

To maintain balance in genetic information the replicated chromosomes of the mother cell divide equally to create two daughter cells, each with a perfectly identical copy of the cellular genetic material. Delivery of chromosomes to each daughter cell during cell division is mediated by the centromere, the fundamental requisite for chromosome inheritance (Figure 1). Indeed, centromeres play a major role in chromosome segregation and cell division driving the assembly of the kinetochore, the protein complex at which the spindle attaches during mitosis and meiosis. Failure in these processes can lead to chromosome mis-segregation and, consequently, to numerical and structural alterations, which in turn can give rise to conditions such as aneuploidy and/or chromosome instability (CIN), both common features of cancer cells.

The Fachinetti lab is interested in understanding how chromosome inheritance is achieved with such high fidelity by identifying how centromeres are established, the mechanisms that drive centromere function, how their integrity is preserved across the cell cycle and the role that centromere failure plays in genome stability.

Figure 2: Overview of the strategies/techniques used to determine the mechanisms of chromosome segregation and integrity including genome editing, cell biology, cytogenetic and genome-wide techniques.
Figure 2: Overview of the strategies/techniques used to determine the mechanisms of chromosome segregation and integrity including genome editing, cell biology, cytogenetic and genome-wide techniques.

Although natural human centromeres are positioned within specific megabase chromosomal regions containing α-satellite DNA repeats, the centromere position is epigenetically specified (Nechemia-Arbely*, Fachinetti*, et al, ECR, 2012). Using gene targeting in human cells and fission yeast, we have previously demonstrated that chromatin containing the centromere-specific histone H3 variant CENP-A is the essential epigenetic mark that acts through a conserved two-step mechanism to identify, maintain and propagate centromere function indefinitely (Fachinetti et al., NCB, 2013). Additionally, we have established that centromere function is not all epigenetically derived; the binding of CENP-B, the only known mammalian centromeric DNA sequence-specific binding protein (present in all human centromeres except the Y chromosome), enhances the fidelity of human centromere function by reinforcing kinetochore formation (Fachinetti et al., Dev Cell, 2015). Our research program is based on an integrated approach that combines the use of engineered cell culture models to conditionally control the stability of endogenous proteins [with an auxin-inducible degron (AID) (Holland*, Fachinetti* et al., PNAS, 2012)] with cell imaging, cytogenetic analysis, proteomic approaches and genome-wide and single molecule technologies (Figure 2). In particular, we are currently studying the molecular mechanism for centromere identification and the importance of DNA sequences for the maintenance of centromere function and integrity. Additional work in the lab is aimed towards the characterization of components that are essential for successful kinetochore nucleation.

Key publications

Year of publication 2017

Fachinetti D, Logsdon GA, Abdullah A, Selzer EB, Cleveland DW, Black BE (2017 Jan 9)

CENP-A Modifications on Ser68 and Lys124 Are Dispensable for Establishment, Maintenance, and Long-Term Function of Human Centromeres.

Dev Cell : 40 : 104-113 : DOI : 10.1016/j.devcel.2016.12.014

Year of publication 2016

Sebastian Hoffmann, Marie Dumont, Viviana Barra, Peter Ly, Yael Nechemia-Arbely, Moira A McMahon, Solène Hervé, Don W Cleveland, Daniele Fachinetti (2016 Nov 24)

CENP-A Is Dispensable for Mitotic Centromere Function after Initial Centromere/Kinetochore Assembly.

Cell reports : 2394-2404 : DOI : 10.1016/j.celrep.2016.10.084

Year of publication 2015

Daniele Fachinetti, Joo Seok Han, Moira A McMahon, Peter Ly, Amira Abdullah, Alex J Wong, Don W Cleveland (2015 May 4)

DNA Sequence-Specific Binding of CENP-B Enhances the Fidelity of Human Centromere Function.

Developmental cell : 314-27 : DOI : 10.1016/j.devcel.2015.03.020

Year of publication 2013

Daniele Fachinetti, H Diego Folco, Yael Nechemia-Arbely, Luis P Valente, Kristen Nguyen, Alex J Wong, Quan Zhu, Andrew J Holland, Arshad Desai, Lars E T Jansen, Don W Cleveland (2013 Feb 7)

A two-step mechanism for epigenetic specification of centromere identity and function.

Nature cell biology : 1056-66 : DOI : 10.1038/ncb2805

Year of publication 2012

Andrew J Holland, Daniele Fachinetti, Joo Seok Han, Don W Cleveland (2012 Nov 12)

Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America : E3350-7 : DOI : 10.1073/pnas.1216880109
Yael Nechemia-Arbely, Daniele Fachinetti, Don W Cleveland (2012 Mar 23)

Replicating centromeric chromatin: spatial and temporal control of CENP-A assembly.

Experimental cell research : 1353-60 : DOI : 10.1016/j.yexcr.2012.04.007
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