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| Status |
Public on May 24, 2022 |
| Title |
DREAM Interrupted: Severing LIN-35-MuvB association in Caenorhabditis elegans impairs DREAM function but not its chromatin localization |
| Organism |
Caenorhabditis elegans |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
The highly conserved DREAM transcriptional repressor complex contains an RB-like pocket protein, an E2F-DP transcription factor heterodimer, and the 5-subunit MuvB complex. Using CRISPR/Cas9 targeted mutagenesis, we disrupted the interaction between the sole Caenorhabditis elegans pocket protein LIN-35 and the MuvB subunit LIN-52. A triple alanine substitution of LIN-52's LxCxE motif (3A) severed LIN-35-MuvB association and caused classical DREAM mutant phenotypes, including synthetic multiple vulvae, high-temperature arrest, and ectopic expression of germline genes in the soma. We performed RNA-seq in lin-52(3A) mutant late embryos (4 replicates) compared to lin-52(WT) wild-type late embryos (4 replicates) to assess the genome-wide effects on gene expression that result from severing LIN-35-MuvB association.
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| Overall design |
We examined 4 biological replicates each of RNA samples (8 total samples) from lin-52(bn139[lin-52::GFP::3xFLAG]) (WT) and lin-52(bn151(lin-52[L42A,C44A,E46A]::GFP::3xFLAG)) (3A) late embryos. Gene expression was compared between 3A and WT to identify misexpressed genes following loss of LIN-35-MuvB association.
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| Contributor(s) |
Goetsch PD, Strome S |
| Citation(s) |
35554539 |
| |
| Submission date |
Mar 23, 2022 |
| Last update date |
May 27, 2022 |
| Contact name |
Paul Donald Goetsch |
| E-mail(s) |
[email protected]
|
| Phone |
906487179
|
| Organization name |
Michigan Technological University
|
| Department |
Biological Sciences
|
| Street address |
1400 Townsend Drive
|
| City |
Houghton |
| State/province |
MI |
| ZIP/Postal code |
49931 |
| Country |
USA |
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| Platforms (1) |
| GPL25147 |
HiSeq X Ten (Caenorhabditis elegans) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA819162 |
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