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The microarchitecture of DNA replication domains
Philimonenko A A, Hodný Z, Jackson D A, Hozák P
Jazyk angličtina Země Německo
NLK
SpringerLink Journals
od 2005-01-01 do 2008-12-31
ProQuest Central
od 1997-01-01 do Před 1 rokem
Medline Complete (EBSCOhost)
od 2000-01-01 do Před 1 rokem
Nursing & Allied Health Database (ProQuest)
od 1997-01-01 do Před 1 rokem
Health & Medicine (ProQuest)
od 1997-01-01 do Před 1 rokem
Public Health Database (ProQuest)
od 1997-01-01 do Před 1 rokem
- MeSH
- buněčné jádro metabolismus ultrastruktura MeSH
- cyklin B genetika MeSH
- DNA-polymerasa I metabolismus MeSH
- DNA genetika chemie MeSH
- financování organizované MeSH
- HeLa buňky MeSH
- imunoelektronová mikroskopie MeSH
- imunohistochemie MeSH
- lamin typ B genetika MeSH
- lidé MeSH
- monoklonální protilátky diagnostické užití MeSH
- proteinkinasa CDC2 genetika MeSH
- proteiny buněčného cyklu fyziologie MeSH
- replikace DNA genetika MeSH
- RNA biosyntéza metabolismus MeSH
- zalévání tkání MeSH
- Check Tag
- lidé MeSH
Most DNA synthesis in HeLa cell nucleus is concentrated in discrete foci. These synthetic sites can be identified by electron microscopy after allowing permeabilized cells to elongate nascent DNA in the presence of biotin-dUTP. Biotin incorporated into nascent DNA can be then immunolabeled with gold particles. Two types of DNA synthetic sites/replication factories can be distinguished at ultrastructural level: (1) electron-dense structures--replication bodies (RB), and (2) focal replication sites with no distinct underlying structure--replication foci (RF). The protein composition of these synthetic sites was studied using double immunogold labeling. We have found that both structures contain (a) proteins involved in DNA replication (DNA polymerase alpha, PCNA), (b) regulators of the cell cycle (cyclin A, cdk2), and (c) RNA processing components like Sm and SS-B/La auto antigens, p80-coilin, hnRNPs A1 and C1/C2. However, at least four regulatory and structural proteins (Cdk1, cyclin B1, PML and lamin B1) differ in their presence in RB and RF. Moreover, in contrast to RF, RB have structural organization. For example, while DNA polymerase alpha, PCNA and hnRNP A1 were diffusely spread throughout RB, hnRNP C1/C2 was found only at the very outside. Surprisingly, RB contained only small amounts of DNA. In conclusion, synthetic sites of both types contain similar but not the same sets of proteins. RB, however, have more developed microarchitecture, apparently with specific functional zones. This data suggest possible differences in genome regions replicated by these two types of replication factories.
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