TY - JOUR
T1 - A dual transcriptional reporter and CDK-activity sensor marks cell cycle entry and progression in C. elegans
AU - Van Rijnberk, Lotte M.
AU - Van Der Horst, Suzanne E M
AU - Van Den Heuvel, Sander
AU - Ruijtenberg, Suzan
N1 - Funding Information:
We thank R. Korswagen and M. Boxem for strains, and members of the van den Heuvel and Boxem groups for help and discussion. We thank Eugene Katrukah (A. Akhmanova lab) for help and expertise with analyzing fluorescence intensities and calculating cytoplasmic over nuclear ratios. Some strains were provided by the CGC, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440).
Publisher Copyright:
© 2017 van Rijnberk 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.
PY - 2017/2/1
Y1 - 2017/2/1
N2 - Development, tissue homeostasis and tumor suppression depend critically on the correct regulation of cell division. Central in the cell division process is the decision whether to enter the next cell cycle and commit to going through the S and M phases, or to remain temporarily or permanently arrested. Cell cycle studies in genetic model systems could greatly benefit from visualizing cell cycle commitment in individual cells without the need of fixation. Here, we report the development and characterization of a reporter to monitor cell cycle entry in the nematode C. elegans. This reporter combines the mcm-4 promoter, to reveal Rb/E2F-mediated transcriptional control, and a live-cell sensor for CDK-activity. The CDK sensor was recently developed for use in human cells and consists of a DNA Helicase fragment fused to eGFP. Upon phosphorylation by CDKs, this fusion protein changes in localization from the nucleus to the cytoplasm. The combined regulation of transcription and subcellular localization enabled us to visualize the moment of cell cycle entry in dividing seam cells during C. elegans larval development. This reporter is the first to reflect cell cycle commitment in C. elegans and will help further genetic studies of the mechanisms that underlie cell cycle entry and exit.
AB - Development, tissue homeostasis and tumor suppression depend critically on the correct regulation of cell division. Central in the cell division process is the decision whether to enter the next cell cycle and commit to going through the S and M phases, or to remain temporarily or permanently arrested. Cell cycle studies in genetic model systems could greatly benefit from visualizing cell cycle commitment in individual cells without the need of fixation. Here, we report the development and characterization of a reporter to monitor cell cycle entry in the nematode C. elegans. This reporter combines the mcm-4 promoter, to reveal Rb/E2F-mediated transcriptional control, and a live-cell sensor for CDK-activity. The CDK sensor was recently developed for use in human cells and consists of a DNA Helicase fragment fused to eGFP. Upon phosphorylation by CDKs, this fusion protein changes in localization from the nucleus to the cytoplasm. The combined regulation of transcription and subcellular localization enabled us to visualize the moment of cell cycle entry in dividing seam cells during C. elegans larval development. This reporter is the first to reflect cell cycle commitment in C. elegans and will help further genetic studies of the mechanisms that underlie cell cycle entry and exit.
UR - https://www.scopus.com/pages/publications/85011588234
U2 - 10.1371/journal.pone.0171600
DO - 10.1371/journal.pone.0171600
M3 - Article
C2 - 28158315
AN - SCOPUS:85011588234
SN - 1932-6203
VL - 12
JO - PLoS ONE [E]
JF - PLoS ONE [E]
IS - 2
M1 - e0171600
ER -