PXD011113-1

PXD011113 is an original dataset announced via ProteomeXchange.

Title	De novo reconstitution and isolation of nucleosomes using the wheat germ cell-free protein synthesis
Description	DNA is packaged with histones and non-histone proteins to form chromatin that impinges on all nuclear processes such as transcription, replication and repair. A complete molecular understanding of these processes requires analysis of chromatin context. In this study, we report a simple and robust system using the wheat cell-free protein synthesis to assemble chromatin. We report two assembly protocols, the post-translational nucleosome assembly (PNAP) and co-translational nucleosome assembly (CNAP), both of which resulted in a native and unmodified form of chromatin. In the PNAP, a mixture of four unpurified core histones and chromatin assembly factors was combined with other components and incubated with DNA. In the CNAP, mRNAs encoding for these proteins were incubated with DNA in the translation mixture, and concomitantly assembled chromatin. Furthermore, we developed a rapid one-step method to isolate the assembled chromatin using divalent cations. Isolated chromatin was assessed for the stoichiometry of histones, nucleosome arrays, supercoiling, and as a substrate for histone modification enzymes. Our methods provide a new avenue to assemble customized chromatin to understand chromatin structure and function.
HostingRepository	PanoramaPublic
AnnounceDate	2026-06-17
AnnouncementXML	Submission_2026-06-17_14:40:21.892.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Nobuaki Takemori
SpeciesList	scientific name: Drosophila melanogaster; NCBI TaxID: 7227;
ModificationList	Deamidated
Instrument	QTRAP 5500

Revision	Datetime	Status	ChangeLog Entry
0	2018-09-17 21:54:37	ID requested
⏵ 1	2026-06-17 14:40:23	announced

Endo Y, Takemori N, Nagy SK, Okimune KI, Kamakaka R, Onouchi H, Takasuka TE, De novo reconstitution of chromatin using wheat germ cell-free protein synthesis. FEBS Open Bio, 11(6):1552-1564(2021) [pubmed]

submitter keyword: Wheat germ cell-free protein synthesis

Nobuaki Takemori
contact affiliation	Ehime University
contact email	takemori@m.ehime-u.ac.jp
lab head
Nobuaki Takemori
contact affiliation	Ehime University
contact email	takemori@m.ehime-u.ac.jp
dataset submitter

Panorama Public dataset URI