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Status |
Public on Jul 22, 2020 |
Title |
Repression of p53-target gene Bbc3/puma by MYSM1 is essential for the survival of hematopoietic multipotent progenitors and contributes to stem cell maintenance |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
MYSM1 is a transcriptional regulator essential for HSC function and hematopoiesis. We established that p53 activation is a common mechanism mediating HSC dysfunction, MPP depletion, and lymphopenia in Mysm1-deficiency, however, the specific p53-induced effectors that trigger dysfunction in Mysm1-deficient HSPCs remain unknown. Here, we performed RNA-Seq of Lin-cKit+Sca1+CD150+ and CD150- HSPCs from Mysm1-/-Puma-/-, Mysm1-/-Puma+/-, Puma-/-, and wild-type mice; (Mysm1-/-Puma+/- phenocopies Mysm1-/-). We showed that Bbc3/PUMA is the primary non-redundant mediator of MPP depletion in Mysm1-deficiency and contributes to HSC dysfunction, whereas depletion of lymphoid-lineage cells involves PUMA-independent p53 activities. We also identified a broad downregulation of genes encoding protein components of the ribosome (RP-genes) and other regulators of translation in Mysm1-deficiency, and the downregulation persisted in Mysm1-/-Puma-/-.
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Overall design |
We performed RNA-Seq transcriptional profiling of FACS-sorted primary hematopoietic stem and progenitor cells. Briefly, bone marrow cells were harvested from Mysm1-/-Puma-/-, Mysm1-/-Puma+/-, Puma-/-, and wild-type mice; (Mysm1-/-Puma+/- phenocopies Mysm1-/-), and sorted using FACS into primary HSPCs, gating on Lin-cKit+Sca1+CD150+ and CD150-. RNA was isolated from the FACS-sorted cells using the MagMAX total RNA kit (Ambion, Life Technologies), and quality assessed using Bioanalyzer RNA Pico chips (Agilent). RNA (10 ng) was rRNA-depleted and used for library preparation with the SMARTer Stranded RNA-Seq kit (Takara Clontech, Mountain View, CA, USA) designed for low input. The RNA-seq libraries were sequenced on an Illumina HiSeq 2000 sequencer in a paired-end 50-bp configuration.
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Contributor(s) |
Wang H, Belle J, Petrov J, Langlais D, Nijnik A |
Citation(s) |
26768662, 34114734 |
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Submission date |
May 15, 2020 |
Last update date |
Jul 20, 2021 |
Contact name |
David Langlais |
E-mail(s) |
david.langlais@mcgill.ca
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Organization name |
McGill University
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Department |
Human Genetics
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Lab |
Inflammation Genomics Lab
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Street address |
740 Ave Dr Penfield, rm 4203, McGill Genome Centre
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City |
MONTREAL |
State/province |
QC |
ZIP/Postal code |
H3A0G1 |
Country |
Canada |
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Platforms (1) |
GPL13112 |
Illumina HiSeq 2000 (Mus musculus) |
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Samples (29)
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This SubSeries is part of SuperSeries: |
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Relations |
BioProject |
PRJNA633124 |
SRA |
SRP261873 |
Supplementary file |
Size |
Download |
File type/resource |
GSE150665_Mysm1Puma_Normalized_CPM_matrix.txt.gz |
2.0 Mb |
(ftp)(http) |
TXT |
GSE150665_RAW.tar |
5.2 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
Processed data are available on Series record |
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