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Differential Requirements for MCM Proteins in DNA Replication in Drosophila S2 Cells.

Crevel, G; Hashimoto, R; Vass, S; Sherkow, J; Yamaguchi, M; Heck, MM; Cotterill, S (2007) Differential Requirements for MCM Proteins in DNA Replication in Drosophila S2 Cells. PLOS ONE, 2 (9). e833. ISSN 1932-6203 https://doi.org/10.1371/journal.pone.0000833
SGUL Authors: Cotterill, Susan Margaret

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Abstract

BACKGROUND: The MCM2-7 proteins are crucial components of the pre replication complex (preRC) in eukaryotes. Since they are significantly more abundant than other preRC components, we were interested in determining whether the entire cellular content was necessary for DNA replication in vivo. METHODOLOGY/PRINCIPLE FINDINGS: We performed a systematic depletion of the MCM proteins in Drosophila S2 cells using dsRNA-interference. Reducing MCM2-6 levels by >95-99% had no significant effect on cell cycle distribution or viability. Depletion of MCM7 however caused an S-phase arrest. MCM2-7 depletion produced no change in the number of replication forks as measured by PCNA loading. We also depleted MCM8. This caused a 30% reduction in fork number, but no significant effect on cell cycle distribution or viability. No additive effects were observed by co-depleting MCM8 and MCM5. CONCLUSIONS/SIGNIFICANCE: These studies suggest that, in agreement with what has previously been observed for Xenopus in vitro, not all of the cellular content of MCM2-6 proteins is needed for normal cell cycling. They also reveal an unexpected unique role for MCM7. Finally they suggest that MCM8 has a role in DNA replication in S2 cells.

Item Type: Article
Additional Information: ©2007 Crevel et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Keywords: Animals, Cell Line, DNA Replication, Drosophila, Drosophila Proteins, Mutation, RNA Interference, Science & Technology, Multidisciplinary Sciences, Science & Technology - Other Topics
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Academic Structure > Molecular and Clinical Sciences Research Institute (MCS) > Cell Sciences (INCCCS)
Journal or Publication Title: PLOS ONE
ISSN: 1932-6203
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Dates:
DateEvent
5 September 2007Published
Web of Science ID: WOS:000207455500017
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URI: http://openaccess.sgul.ac.uk/id/eprint/2460
Publisher's version: https://doi.org/10.1371/journal.pone.0000833

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