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Evidence for reciliation of RPE1 cells in late G1 phase, and ciliary localisation of cyclin B1

Evidence for reciliation of RPE1 cells in late G1 phase, and ciliary localisation of cyclin B1
Evidence for reciliation of RPE1 cells in late G1 phase, and ciliary localisation of cyclin B1
The primary cilium, an organelle that transduces extracellular signals important for development and tissue homeostasis, is typically assembled upon cell cycle exit and disassembled upon cell cycle re-entry. Cilium assembly is thought to be suppressed in cycling cells, however the extent of suppression is not clear. For example, primary cilia are present in certain proliferating cells during development, and a period of reciliation has been reported to occur in late G1 in murine 3T3 cells released from serum starvation-induced quiescence. Human retinal pigmented epithelial (hTERT-RPE1; herein, RPE1) cells are commonly used to investigate pathways regulating cilium disassembly, however the ciliary disassembly profile of these cells remains uncertain. A period of reciliation has not been observed. Here, we analyse the ciliary disassembly profile of RPE1 cells by immunofluorescence microscopy. The results suggest a profile similar to 3T3 cells, including a period of reciliation in late G1 and a second wave of deciliation in S phase. We present evidence that arresting cells in early S phase with hydroxyurea or excess thymidine prevents the second wave of deciliation, and that deciliation is initiated shortly after release from a thymidine block, consistent with coupling to DNA replication. These findings support the often overlooked notion that cilium formation can occur in late G1, and suggest that RPE1 cells could serve as a model system for studying the molecular pathways that direct this process, in addition to those that stimulate cilium disassembly. We also present immunofluorescence data indicating that cyclin B1 localises to primary cilia.
2211-5463
334-340
Spalluto, Cosma
6802ad50-bc38-404f-9a19-40916425183b
Wilson, David I.
1500fca1-7082-4271-95f4-691f1d1252a2
Hearn, Tom
2665cc10-6632-47cb-9460-bd0ea745380e
Spalluto, Cosma
6802ad50-bc38-404f-9a19-40916425183b
Wilson, David I.
1500fca1-7082-4271-95f4-691f1d1252a2
Hearn, Tom
2665cc10-6632-47cb-9460-bd0ea745380e

Spalluto, Cosma, Wilson, David I. and Hearn, Tom (2013) Evidence for reciliation of RPE1 cells in late G1 phase, and ciliary localisation of cyclin B1. FEBS Open Bio, 3, 334-340. (doi:10.1016/j.fob.2013.08.002). (PMID:24251092)

Record type: Article

Abstract

The primary cilium, an organelle that transduces extracellular signals important for development and tissue homeostasis, is typically assembled upon cell cycle exit and disassembled upon cell cycle re-entry. Cilium assembly is thought to be suppressed in cycling cells, however the extent of suppression is not clear. For example, primary cilia are present in certain proliferating cells during development, and a period of reciliation has been reported to occur in late G1 in murine 3T3 cells released from serum starvation-induced quiescence. Human retinal pigmented epithelial (hTERT-RPE1; herein, RPE1) cells are commonly used to investigate pathways regulating cilium disassembly, however the ciliary disassembly profile of these cells remains uncertain. A period of reciliation has not been observed. Here, we analyse the ciliary disassembly profile of RPE1 cells by immunofluorescence microscopy. The results suggest a profile similar to 3T3 cells, including a period of reciliation in late G1 and a second wave of deciliation in S phase. We present evidence that arresting cells in early S phase with hydroxyurea or excess thymidine prevents the second wave of deciliation, and that deciliation is initiated shortly after release from a thymidine block, consistent with coupling to DNA replication. These findings support the often overlooked notion that cilium formation can occur in late G1, and suggest that RPE1 cells could serve as a model system for studying the molecular pathways that direct this process, in addition to those that stimulate cilium disassembly. We also present immunofluorescence data indicating that cyclin B1 localises to primary cilia.

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More information

Published date: 11 August 2013
Organisations: Human Development & Health, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 360078
URI: http://eprints.soton.ac.uk/id/eprint/360078
ISSN: 2211-5463
PURE UUID: d5dcce22-e322-4001-9177-7d72f41f6e74
ORCID for Cosma Spalluto: ORCID iD orcid.org/0000-0001-7273-0844

Catalogue record

Date deposited: 25 Nov 2013 13:06
Last modified: 14 Mar 2023 02:38

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Contributors

Author: Cosma Spalluto ORCID iD
Author: David I. Wilson
Author: Tom Hearn

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