C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2016-10-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/18370 |
| _version_ | 1811181032590802944 |
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| author | ChangHwan Lee Erika B Sorensen Tina R Lynch Judith Kimble |
| author_facet | ChangHwan Lee Erika B Sorensen Tina R Lynch Judith Kimble |
| author_sort | ChangHwan Lee |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:12:33Z |
| format | Article |
| id | doaj.art-56590abae95c4d91bc2ce30764ada7f1 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:12:33Z |
| publishDate | 2016-10-01 |
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| spelling | doaj.art-56590abae95c4d91bc2ce30764ada7f12022-12-22T04:32:28ZengeLife Sciences Publications LtdeLife2050-084X2016-10-01510.7554/eLife.18370C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell poolChangHwan Lee0Erika B Sorensen1Tina R Lynch2Judith Kimble3https://orcid.org/0000-0001-5622-2073Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, United States; Department of Biochemistry, University of Wisconsin-Madison, Madison, United StatesHoward Hughes Medical Institute, University of Wisconsin-Madison, Madison, United States; Department of Biochemistry, University of Wisconsin-Madison, Madison, United StatesDepartment of Biochemistry, University of Wisconsin-Madison, Madison, United StatesHoward Hughes Medical Institute, University of Wisconsin-Madison, Madison, United States; Department of Biochemistry, University of Wisconsin-Madison, Madison, United StatesC. 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.https://elifesciences.org/articles/18370active transcription sitesingle-molecule FISHsygl-1lst-1 |
| spellingShingle | ChangHwan Lee Erika B Sorensen Tina R Lynch Judith Kimble C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool eLife active transcription site single-molecule FISH sygl-1 lst-1 |
| title | C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool |
| title_full | C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool |
| title_fullStr | C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool |
| title_full_unstemmed | C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool |
| title_short | C. elegans GLP-1/Notch activates transcription in a probability gradient across the germline stem cell pool |
| title_sort | c elegans glp 1 notch activates transcription in a probability gradient across the germline stem cell pool |
| topic | active transcription site single-molecule FISH sygl-1 lst-1 |
| url | https://elifesciences.org/articles/18370 |
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