We have analyzed human centromeric chromatin containing CENPA from HeLa cells in culture using Chromatin IP followed by atomic force microscopy measurements and mass spectrophotometry. Thus far, we have identified that the CENPA chromatin structure is uniquely organized during interphase, and has fundamentally different characteristics from the rest of the chromatin which packages the genome. We plan to extend our analysis using live imaging coupled with AFM across the cell cycle to address how CENP-A is assembled and retained, as well as how it participates in non-centromeric functions at ectopic loci. We have discovered that CENP-A changes its structure over the cell cycle. This manuscript has been submitted for publication.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC011207-03
Application #
8349396
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2011
Total Cost
$299,919
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Nuccio, A G; Bui, M; Dalal, Y et al. (2017) Mass Spectrometry-Based Methodology for Identification of Native Histone Variant Modifications From Mammalian Tissues and Solid Tumors. Methods Enzymol 586:275-290
Vlijm, R; Kim, S H; De Zwart, P L et al. (2017) The supercoiling state of DNA determines the handedness of both H3 and CENP-A nucleosomes. Nanoscale 9:1862-1870
Bui, Minh; Pitman, Mary; Nuccio, Arthur et al. (2017) Internal modifications in the CENP-A nucleosome modulate centromeric dynamics. Epigenetics Chromatin 10:17
Zhao, Haiqing; Winogradoff, David; Bui, Minh et al. (2016) Promiscuous Histone Mis-Assembly Is Actively Prevented by Chaperones. J Am Chem Soc :
Kim, Sung Hyun; Vlijm, Rifka; van der Torre, Jaco et al. (2016) CENP-A and H3 Nucleosomes Display a Similar Stability to Force-Mediated Disassembly. PLoS One 11:e0165078
Quénet, D; Sturgill, D; Dalal, Y (2016) Identifying Centromeric RNAs Involved in Histone Dynamics In Vivo. Methods Enzymol 573:445-66
Quénet, Delphine; Dalal, Yamini (2015) Correction: a long non-coding RNA is required for targeting centromeric protein A to the human centromere. Elife 4:
Winogradoff, David; Zhao, Haiqing; Dalal, Yamini et al. (2015) Shearing of the CENP-A dimerization interface mediates plasticity in the octameric centromeric nucleosome. Sci Rep 5:17038
Quénet, Delphine; Dalal, Yamini (2014) A long non-coding RNA is required for targeting centromeric protein A to the human centromere. Elife 3:e03254
Walkiewicz, Marcin P; Dimitriadis, Emilios K; Dalal, Yamini (2014) CENP-A octamers do not confer a reduction in nucleosome height by AFM. Nat Struct Mol Biol 21:2-3

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