Options
Cell Cycle-Dependent Mobility of Cdc45 Determined in vivo by Fluorescence Correlation Spectroscopy
Date Issued
2012-04-19
Date Available
2013-11-28T14:50:40Z
Abstract
Eukaryotic DNA replication is a dynamic process requiring the co-operation of specific replication proteins. We measured the mobility of eGFP-Cdc45 by Fluorescence Correlation Spectroscopy (FCS) in vivo in asynchronous cells and in cells synchronized at the G1/S transition and during S phase. Our data show that eGFP-Cdc45 mobility is faster in G1/S transition compared to S phase suggesting that Cdc45 is part of larger protein complex formed in S phase. Furthermore, the size of complexes containing Cdc45 was estimated in asynchronous, G1/S and S phase-synchronized cells using gel filtration chromatography; these findings complemented the in vivo FCS data. Analysis of the mobility of eGFP-Cdc45 and the size of complexes containing Cdc45 and eGFP-Cdc45 after UVC-mediated DNA damage revealed no significant changes in diffusion rates and complex sizes using FCS and gel filtration chromatography analyses. This suggests that after UV-damage, Cdc45 is still present in a large multi-protein complex and that its mobility within living cells is consistently similar following UVC-mediated DNA damage.
Other Sponsorship
SFI
Type of Material
Journal Article
Publisher
Public Library of Science
Journal
PLoS ONE
Volume
7
Issue
4
Copyright (Published Version)
2012 Public Library of Science
Language
English
Status of Item
Peer reviewed
This item is made available under a Creative Commons License
File(s)
Owning collection
Scopus© citations
14
Acquisition Date
Apr 17, 2024
Apr 17, 2024
Views
1364
Last Month
1
1
Acquisition Date
Apr 17, 2024
Apr 17, 2024
Downloads
319
Acquisition Date
Apr 17, 2024
Apr 17, 2024