C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool
- Howard Hughes Medical Institute, University of Wisconsin-Madison, United States
- Wabash College, United States
- University of Wisconsin-Madison, United States
Abstract
C. elegans Notch signaling maintains a pool of germline stem cells within their single-celled mesenchymal niche. Here we investigate the Notch transcriptional response in germline stem cells using single-molecule fluorescence in situ hybridization coupled with automated, high-throughput quantitation. This approach allows us to distinguish Notch-dependent nascent transcripts in the nucleus from mature mRNAs in the cytoplasm. We find that Notch-dependent active transcription sites occur in a probabilistic fashion and, unexpectedly, do so in a steep gradient across the stem cell pool. Yet these graded nuclear sites create a nearly uniform field of mRNAs that extends beyond the region of transcriptional activation. Therefore, active transcription sites provide a precise view of where the Notch-dependent transcriptional complex is productively engaged. Our findings offer a new window into the Notch transcriptional response and demonstrate the importance of assaying nascent transcripts at active transcription sites as a readout for canonical signaling.
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Author details
Funding
American Cancer Society (PF-14-147-01-DDC)
- Erika B Sorensen
Howard Hughes Medical Institute
- Judith Kimble
American Cancer Society (PF-14-147-01-DDC)
- Erika B Sorensen
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2016, Lee et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool
eLife 5:e18370.
https://doi.org/10.7554/eLife.18370
