A dual transcriptional reporter and CDK-activity sensor marks cell cycle entry and progression in C. elegans.
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.
Developmental Biology, Department of Biology, Faculty of Science, Utrecht University Padualaan 8, CH Utrecht, The Netherlands.
Lotte M
LM
van Rijnberk
Developmental Biology, Department of Biology, Faculty of Science, Utrecht University Padualaan 8, CH Utrecht, The Netherlands.
Suzanne E M
SE
van der Horst
Developmental Biology, Department of Biology, Faculty of Science, Utrecht University Padualaan 8, CH Utrecht, The Netherlands.
Sander
S
van den Heuvel
Developmental Biology, Department of Biology, Faculty of Science, Utrecht University Padualaan 8, CH Utrecht, The Netherlands.
Suzan
S
Ruijtenberg
D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics
D029742:Caenorhabditis elegans Proteins; D018844:Cyclin-Dependent Kinases