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Series GSE55949 Query DataSets for GSE55949
Status Public on Oct 01, 2014
Title Histone H3.3 and its proteolytically processed form drive a cellular senescence program
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary In this study, we have uncovered novel proteolytic processing of the histone H3 tail in senescence models in primary fibroblasts and melanocytes. Cleavage of H3 tail occurs at two distinct residues and is mediated by Cathepsin L. We show that variant H3.3 is preferentially cleaved, and that cleaved histones are associated with chromatin and incorporated into nucleosomes. We also found that the histone chaperone ASF1a is required for chromatin incorporation of the cleaved histone species. Further, we show that overexpression of cleaved H3.3 induces a senescence program in fibroblasts in the absensence of oncogenic signaling.
Overall design For the RNA-seq studies, growing IMR90 fibroblasts were compared to cells induced to senesce via oncogene activation or cleaved H3.3 overexpression. Growing controls consist of IMR90 cells infected with empty retroviral construct pBabe and grown under normal conditions for 13 days prior to RNA isolation. For oncogene-induced senescence samples, IMR90s carrying a tamoxifen-inducible H-RasV12 retroviral construct were induced to senesce by addition of 10nM tamoxifen to the media for 8 days. Finally, IMR90s were infected with a retroviral construct expressing the cleaved form of H3.3 with a C-terminal Flag tag. RNA samples form this group were isolated at days 3 (early) and 13 (late) post-infection. In all cases, total RNA samples were isolated using RNeasy kit (Qiagen) and prepared at the Icahn School of Medicine at Mount Sinai Genomics Core Facility for poly A library construction and sequencing on IlluminaHiSeq 2500.
Contributor(s) Bernstein E, Duarte LF
Citation(s) 25394905
Submission date Mar 17, 2014
Last update date May 15, 2019
Contact name Zichen Wang
Organization name Icahn School of Medicine at Mount Sinai
Department Pharmacological Sciences
Lab Avi Ma'ayan
Street address 1468 Madison Ave
City New York
State/province NY
ZIP/Postal code 10029
Country USA
Platforms (1)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
Samples (4)
GSM1348980 Growing
GSM1348981 Oncogene-induced senescence
GSM1348982 H3.3cs1-induced senescence early time point
BioProject PRJNA241413
SRA SRP040243

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Supplementary file Size Download File type/resource
GSE55949_RAW.tar 950.0 Kb (http)(custom) TAR (of TXT)
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Raw data are available in SRA
Processed data provided as supplementary file

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