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Series GSE150665 Query DataSets for GSE150665
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
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-/-.
 
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.
 
Contributor(s) Wang H, Belle J, Petrov J, Langlais D, Nijnik A
Citation(s) 26768662, 34114734
Submission date May 15, 2020
Last update date Jul 20, 2021
Contact name David Langlais
E-mail(s) david.langlais@mcgill.ca
Organization name McGill University
Department Human Genetics
Lab Inflammation Genomics Lab
Street address 740 Ave Dr Penfield, rm 4203, McGill Genome Centre
City MONTREAL
State/province QC
ZIP/Postal code H3A0G1
Country Canada
 
Platforms (1)
GPL13112 Illumina HiSeq 2000 (Mus musculus)
Samples (29)
GSM4555943 CD150POS_A1_Mysm1-KO_Puma-HET
GSM4555944 CD150POS_A2_Mysm1-KO_Puma-HET
GSM4555945 CD150POS_A3_Mysm1-KO_Puma-HET
This SubSeries is part of SuperSeries:
GSE150667 MYSM1
Relations
BioProject PRJNA633124
SRA SRP261873

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

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 SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file
Processed data are available on Series record

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