Cell Cycle-Dependent Mobility of Cdc45 Determined in vivo by Fluorescence Correlation Spectroscopy

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Title: Cell Cycle-Dependent Mobility of Cdc45 Determined in vivo by Fluorescence Correlation Spectroscopy
Authors: Broderick, Ronan
Ramadurai, Sivaramakrishnan
Tóth, Katalin
et al.
Permanent link: http://hdl.handle.net/10197/4997
Date: 19-Apr-2012
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.
Type of material: Journal Article
Publisher: Public Library of Science
Copyright (published version): 2012 Public Library of Science
Keywords: eGFP-Cdc45Eukaryotic DNA replicationFluorescence Correlation Spectroscopy (FCS)
DOI: 10.1371/journal.pone.0035537
Language: en
Status of Item: Peer reviewed
Appears in Collections:SBI Research Collection

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