Alterations in chromatin structure and gene expression are a hallmark of cancer. Proteins involved in chromatin remodeling complexes (CRCs) directly affect chromatin structure by modifying DMA and histones, and CRCs targeting is altered in cancer. Structural proteins like NuMA, considered as organizers of nuclear architecture, also interact with chromatin, and their distribution is altered in cancer. However, the role of NuMA in the control of gene expression is unknown. NuMA co-isolates with components of ATP-dependent CRCs. Altering NuMA using an antibody against its C-terminus (CT) or by expressing the distal portion of its CT modifies chromatin structure and the phenotype of breast epithelial cells. CT-truncated NuMA and mutated NuMA are implicated in leukemia and breast cancer development, respectively. We propose to investigate the mechanisms by which NuMA controls chromatin structure and gene expression in breast epithelial cells. The hypothesis is that NuMA participates in the control of gene transcription by interacting with components of CRCs, in order to target these complexes to specific genes.
Three aims are proposed: (1) To identify the specific CRCs in which NuMA is involved by assessing the interaction of NuMA with components of SNF2h and BAF types of ATP-dependent CRCs by affinity chromatography and co- immunoprecipitation, and assessing the effect of inhibiting NuMA expression on the assembly and function of CRCs;(2) To analyze the primary sequence of NuMA with regards to its involvement in chromatin regulation by comparing the effects of interrupting the function of HPC2-like, GAS41-binding, and CH-binding regions of NuMA on chromatin organization, CRC function, and mammary epithelial differentiation;(3) To define the role of NuMA in gene expression control by position effect, by interfering with NuMA function and evaluating the expression status (on or off) of genes that control cell proliferation in relation to their localization and the presence of NuMA-containing CRCs at their promoter. Significance: A current scientific and medical challenge is to unravel the mechanisms that underlie changes in the machinery or regulation of gene expression during cancer development. Understanding these mechanisms will help develop strategies to prevent cancer progression and control tumor behavior. Thus, it is time to emphasize the role of structural nuclear proteins in gene transcription in order to get a complete picture of gene expression control.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA112017-04S1
Application #
8514778
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Mietz, Judy
Project Start
2007-09-26
Project End
2013-01-31
Budget Start
2012-08-01
Budget End
2013-01-31
Support Year
4
Fiscal Year
2012
Total Cost
$29,097
Indirect Cost
$10,203
Name
Purdue University
Department
Other Basic Sciences
Type
Schools of Veterinary Medicine
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Vidi, Pierre-Alexandre; Liu, Jing; Salles, Daniela et al. (2014) NuMA promotes homologous recombination repair by regulating the accumulation of the ISWI ATPase SNF2h at DNA breaks. Nucleic Acids Res 42:6365-79
Vidi, Pierre-Alexandre; Leary, James F; Lelièvre, Sophie A (2013) Building risk-on-a-chip models to improve breast cancer risk assessment and prevention. Integr Biol (Camb) 5:1110-8
Vidi, Pierre-Alexandre; Bissell, Mina J; Lelièvre, Sophie A (2013) Three-dimensional culture of human breast epithelial cells: the how and the why. Methods Mol Biol 945:193-219
Bazzoun, Dana; Lelièvre, Sophie; Talhouk, Rabih (2013) Polarity proteins as regulators of cell junction complexes: implications for breast cancer. Pharmacol Ther 138:418-27
Teegarden, Dorothy; Romieu, Isabelle; Lelièvre, Sophie A (2012) Redefining the impact of nutrition on breast cancer incidence: is epigenetics involved? Nutr Res Rev 25:68-95
Vidi, Pierre-Alexandre; Chandramouly, Gurushankar; Gray, Matthew et al. (2012) Interconnected contribution of tissue morphogenesis and the nuclear protein NuMA to the DNA damage response. J Cell Sci 125:350-61
Yue, Shuhua; Cardenas-Mora, Juan Manuel; Chaboub, Lesley S et al. (2012) Label-free analysis of breast tissue polarity by Raman imaging of lipid phase. Biophys J 102:1215-23
Lelièvre, Sophie A (2010) Tissue polarity-dependent control of mammary epithelial homeostasis and cancer development: an epigenetic perspective. J Mammary Gland Biol Neoplasia 15:49-63
Lelievre, Sophie A (2009) Contributions of extracellular matrix signaling and tissue architecture to nuclear mechanisms and spatial organization of gene expression control. Biochim Biophys Acta 1790:925-35