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| Status |
Public on Jul 27, 2023 |
| Title |
MSL2 cut and run profiles in S2 cell lines |
| Organism |
Drosophila melanogaster |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
MSL2, the DNA-binding subunit of the Drosophila dosage compensation complex, cooperates with the ubiquitous protein CLAMP to bind MSL recognition elements (MREs) on the X chromosome. We explore the nature of the cooperative binding to these GA-rich, composite sequence elements in reconstituted naïve embryonic chromatin. We found that the cooperativity requires physical interaction between both proteins. Remarkably, disruption of this interaction does not lead to indirect, nucleosome-mediated cooperativity as expected, but to competition. The protein interaction apparently not only increases the affinity for composite binding sites, but also locks both proteins in a defined dimeric state that prevents competition. High Affinity Sites of MSL2 on the X chromosome contain variable numbers of MREs. We find that the cooperation between MSL2/CLAMP is not influenced by MRE clustering or arrangement, but happens largely at the level of individual MREs. The sites where MSL2/CLAMP bind strongly in vitro locate to all chromosomes and show little overlap to an expanded set of X-chromosomal MSL2 in vivo binding sites generated by CUT&RUN. Apparently, the intrinsic MSL2/CLAMP cooperativity is limited to a small selection of potential sites in vivo. This restriction must be due to components missing in our reconstitution, such as roX2 lncRNA.
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| Overall design |
Cut and Run profiles of MSL2 in S2 cell lines
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| Contributor(s) |
Eggers N, Gkountromichos F |
| Citation(s) |
37602401 |
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| Submission date |
Apr 04, 2023 |
| Last update date |
Sep 27, 2023 |
| Contact name |
Nikolas Eggers |
| E-mail(s) |
[email protected]
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| Organization name |
LMU Munich
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| Department |
Molecular Biology
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| Lab |
Becker
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| Street address |
Grosshaderner Strasse 9
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| City |
Munich |
| ZIP/Postal code |
80469 |
| Country |
Germany |
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| Platforms (1) |
| GPL33093 |
NextSeq 1000 (Drosophila melanogaster) |
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| Samples (5)
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| Relations |
| BioProject |
PRJNA952217 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE228935_msl2_cnr.f10.peaks.bed.gz |
8.8 Kb |
(ftp)(http) |
BED |
| GSE228935_msl2_cnr.ipnorm.bw |
98.9 Mb |
(ftp)(http) |
BW |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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