PMID- 26075356
OWN - NLM
STAT- MEDLINE
DCOM- 20151022
LR  - 20181113
IS  - 1476-4679 (Electronic)
IS  - 1465-7392 (Linking)
VI  - 17
IP  - 8
DP  - 2015 Aug
TI  - Reconstitution of mitotic chromatids with a minimum set of purified factors.
PG  - 1014-23
LID - 10.1038/ncb3187 [doi]
AB  - The assembly of mitotic chromosomes, each composed of a pair of rod-shaped 
      chromatids, is an essential prerequisite for accurate transmission of the genome 
      during cell division. It remains poorly understood, however, how this fundamental 
      process might be achieved and regulated in the cell. Here we report an in vitro 
      system in which mitotic chromatids can be reconstituted by mixing a simple 
      substrate with only six purified factors: core histones, three histone chaperones 
      (nucleoplasmin, Nap1 and FACT), topoisomerase II (topo II) and condensin I. We 
      find that octameric nucleosomes containing the embryonic variant H2A.X-F are 
      highly susceptible to FACT and function as the most productive substrate for 
      subsequent actions of topo II and condensin I. Cdk1 phosphorylation of condensin 
      I is the sole mitosis-specific modification required for chromatid 
      reconstitution. This experimental system will enhance our understanding of the 
      mechanisms of action of individual factors and their cooperation during this 
      process.
FAU - Shintomi, Keishi
AU  - Shintomi K
AD  - Chromosome Dynamics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan.
FAU - Takahashi, Tatsuro S
AU  - Takahashi TS
AD  - Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
FAU - Hirano, Tatsuya
AU  - Hirano T
AD  - Chromosome Dynamics Laboratory, RIKEN, Wako, Saitama 351-0198, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150615
PL  - England
TA  - Nat Cell Biol
JT  - Nature cell biology
JID - 100890575
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (High Mobility Group Proteins)
RN  - 0 (Histones)
RN  - 0 (Molecular Chaperones)
RN  - 0 (Multiprotein Complexes)
RN  - 0 (Nucleoplasmins)
RN  - 0 (Nucleosomes)
RN  - 0 (SSRP1 protein, human)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 0 (Transcriptional Elongation Factors)
RN  - 0 (Xenopus Proteins)
RN  - 0 (condensin complexes)
RN  - EC 2.7.11.22 (CDC2 Protein Kinase)
RN  - EC 3.6.1.- (Adenosine Triphosphatases)
RN  - EC 5.99.1.3 (DNA Topoisomerases, Type II)
SB  - IM
CIN - Nat Cell Biol. 2015 Aug;17(8):964-5. doi: 10.1038/ncb3212. PMID: 26239527
CIN - Curr Biol. 2015 Aug 3;25(15):R663-6. doi: 10.1016/j.cub.2015.06.026. PMID: 
      26241143
MH  - Adenosine Triphosphatases/metabolism
MH  - Animals
MH  - CDC2 Protein Kinase/metabolism
MH  - Chromatids/*enzymology
MH  - *Chromatin Assembly and Disassembly
MH  - DNA Topoisomerases, Type II/metabolism
MH  - DNA-Binding Proteins/metabolism
MH  - HeLa Cells
MH  - High Mobility Group Proteins/metabolism
MH  - Histones/genetics/*metabolism
MH  - Humans
MH  - Male
MH  - *Mitosis
MH  - Molecular Chaperones/genetics/*metabolism
MH  - Multiprotein Complexes/metabolism
MH  - Nucleoplasmins/metabolism
MH  - Nucleosomes/enzymology
MH  - Phosphorylation
MH  - Saccharomyces cerevisiae
MH  - Saccharomyces cerevisiae Proteins/genetics/*metabolism
MH  - Spermatozoa/*enzymology
MH  - Transcriptional Elongation Factors/metabolism
MH  - Transfection
MH  - Xenopus Proteins/genetics/*metabolism
MH  - Xenopus laevis
EDAT- 2015/06/16 06:00
MHDA- 2015/10/23 06:00
CRDT- 2015/06/16 06:00
PHST- 2015/01/30 00:00 [received]
PHST- 2015/05/12 00:00 [accepted]
PHST- 2015/06/16 06:00 [entrez]
PHST- 2015/06/16 06:00 [pubmed]
PHST- 2015/10/23 06:00 [medline]
AID - ncb3187 [pii]
AID - 10.1038/ncb3187 [doi]
PST - ppublish
SO  - Nat Cell Biol. 2015 Aug;17(8):1014-23. doi: 10.1038/ncb3187. Epub 2015 Jun 15.