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| Status |
Public on Mar 26, 2024 |
| Title |
DNAJC9 prevents CENP-A mislocalization and chromosomal instability by maintaining the fidelity of H3-H4 supply chains |
| Organism |
Homo sapiens |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
The centromeric histone H3 variant CENP-A is overexpressed in many cancers. Mislocalization of CENP-A to non-centromeric regions contributes to chromosomal instability (CIN), a hallmark of cancer. Despite these observations, pathways that promote and prevent CENP-A mislocalization remain poorly defined. Here, we performed a genome-wide RNAi screen to identify regulators of CENP-A localization. We identified DNAJC9, a J-domain protein as a lead candidate from the screen and showed that cells depleted for DNAJC9 exhibit mislocalization of CENP-A, enrichment of CENP-A in chromatin, and CIN phenotypes. Global interactome analysis showed an enhanced interaction of CENP-A with the replication-associated H3-H4 chaperone MCM2 in DNAJC9-depleted cells and co-depletion of MCM2 and DNAJC9 suppressed CENP-A mislocalization. Furthermore, we showed that cells ablated for the ability of DNAJC9 to promote the proper folding of H3–H4, exhibit CENP-A mislocalization. CUT&RUN Sequencing analysis of genome-wide CENP-A occupancy in DNAJC9-depleted cells identified 16,603 sites of non-centromeric localization, that broadly overlapped with open chromatin regions. Our comprehensive analysis has identified factors that prevent mislocalization of CENP-A and has defined DNAJC9 as an important safeguard that prevents CENP-A mislocalization and CIN.
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| Overall design |
CUT&RUN Sequencing of CENP-A in siNegative and siDNAJC9 transfected hTERT-RPE1-Tet-GFP-CENP-A cells treated with 1 ug/ml DOX for 48 hrs. Two biological repeats per condition were analysed with Mouse anti-CENP-A antibody and Mouse anti-IgG was used as control.
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| Contributor(s) |
Balachandra V, Shrestha RL, Hammond CM, Lin S, Sevilla S, Basrai MA |
| Citation(s) |
38600242 |
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| Submission date |
Jan 16, 2024 |
| Last update date |
Jun 26, 2024 |
| Contact name |
Vinutha Balachandra |
| Organization name |
National Institute for Health
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| Department |
National Cancer Institute, Genetics Branch
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| Lab |
Yeast Genome Stability Section
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| Street address |
10 Center Drive
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Platforms (1) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA1065442 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE253387_RAW.tar |
473.6 Mb |
(http)(custom) |
TAR (of BED, BIGWIG) |
SRA Run Selector |
| Raw data are available in SRA |
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