Chromatin plays two essential, yet by their very nature mutually exclusive roles. On one hand, chromatin is responsible for the extreme degree of DNA compaction that allows the eukaryotic genome to fit into the confines of the nucleus. On the other hand, it must permit the local unraveling of DNA to grant access of the cellular machinery to the genome. Activities that mediate the inter- conversion between various levels of compacted chromatin states are key regulators of all processes requiring access to genomic DNA. Mechanistic and structural insight into these biologically relevant processes is limited. Here, the role of the nucleosome assembly protein 1 (NAP1) family of histone chaperones in maintaining and modulating chromatin structure and fluidity will be investigated.
Aims 1 and 2 address the structural and molecular basis for chaperone interaction with histones, using x-ray diffraction as well as biochemical and in vivo approaches. The mechanism and biological role of NAP1-mediated histone removal and histone exchange will be studied by rigorous kinetic analysis combined with in vivo examination of chromatin structure (aim 3). Finally, the novel hypothesis that a posttranslational modification regulates NAP1 activity in metazoans will be put to test in aim 4. The strength of this proposal lies in a combination of a broad spectrum of structural, biophysical, biochemical and genetic approaches to address the highly significant question of how compacted DNA is made available to the cellular machinery.
The molecular mechanism and biological relevance of histone chaperone-mediated chromatin structure modulation will be investigated, using a combination of structural, biophysical, and genetic approaches.
|Mattiroli, Francesca; Bhattacharyya, Sudipta; Dyer, Pamela N et al. (2017) Structure of histone-based chromatin in Archaea. Science 357:609-612|
|Mattiroli, Francesca; Gu, Yajie; Yadav, Tejas et al. (2017) DNA-mediated association of two histone-bound complexes of yeast Chromatin Assembly Factor-1 (CAF-1) drives tetrasome assembly in the wake of DNA replication. Elife 6:|
|Mattiroli, Francesca; Gu, Yajie; Balsbaugh, Jeremy L et al. (2017) The Cac2 subunit is essential for productive histone binding and nucleosome assembly in CAF-1. Sci Rep 7:46274|
|Pentakota, Satyakrishna; Zhou, Keda; Smith, Charlotte et al. (2017) Decoding the centromeric nucleosome through CENP-N. Elife 6:|
|White, Alison E; Hieb, Aaron R; Luger, Karolin (2016) A quantitative investigation of linker histone interactions with nucleosomes and chromatin. Sci Rep 6:19122|
|Prasad, Rashmi; D'Arcy, Sheena; Hada, Arjan et al. (2016) Coordinated Action of Nap1 and RSC in Disassembly of Tandem Nucleosomes. Mol Cell Biol 36:2262-71|
|Chen, Xu; D'Arcy, Sheena; Radebaugh, Catherine A et al. (2016) Histone Chaperone Nap1 Is a Major Regulator of Histone H2A-H2B Dynamics at the Inducible GAL Locus. Mol Cell Biol 36:1287-96|
|Zhao, Huaying; Ghirlando, Rodolfo; Alfonso, Carlos et al. (2015) A multilaboratory comparison of calibration accuracy and the performance of external references in analytical ultracentrifugation. PLoS One 10:e0126420|
|Dechassa, Mekonnen Lemma; Wyns, Katharina; Luger, Karolin (2014) Scm3 deposits a (Cse4-H4)2 tetramer onto DNA through a Cse4-H4 dimer intermediate. Nucleic Acids Res 42:5532-42|
|Muthurajan, Uma M; Hepler, Maggie R D; Hieb, Aaron R et al. (2014) Automodification switches PARP-1 function from chromatin architectural protein to histone chaperone. Proc Natl Acad Sci U S A 111:12752-7|
Showing the most recent 10 out of 33 publications