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| Status |
Public on Jun 29, 2021 |
| Title |
TENT4A non-canonical poly(A) polymerase regulates DNA-damage tolerance via multiple pathways which are mutated in endometrial cancer |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
TENT4A (PAPD7) is a non-canonical poly(A) polymerase, of which little is known. Here we show that TENT4A is involved in the regulation of multiple biological pathways, and focus on translesion DNA synthesis (TLS), a DNA-damage tolerance process in which unrepaired lesions are bypassed by error-prone DNA polymerases. We show that TENT4A regulates mRNA stability and/or translation of DNA polymerase η, and of the RAD18 E3 ligase that monoubiquitinates PCNA, the latter being a key step in TLS which enables recruitment of error-prone DNA polymerases to damaged DNA sites. Remarkably, in addition to the effect on RAD18 mRNA stability via controlling its poly(A) tail, TENT4A indirectly regulates RAD18 via the tumor suppressor CYLD, and via the long non-coding antisense RNA PAXIP1-AS2, which had no known function. Knocking down the expression of TENT4A or CYLD, or over-expression of PAXIP1-AS2 led each to a decrease in the amount of the RAD18 protein, but the effect on PCNA ubiquitination was variable. Still, TLS was reduced under these conditions, likely due a to a parallel decrease in DNA polymerase η. Bioinformatics analysis revealed that TLS error-prone DNA polymerase genes and their TENT4A-related regulators are frequently mutated in endometrial cancer genomes, suggesting that TLS is dysregulated in endometrial cancer.
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| Overall design |
Human osteosarcoma U2OS cell line and XP12RO (Xeroderma pigmentosum, complementation group A, SV-40 transformed) cells were selected as samples. The shRNA against TENT4A lentiviral particles was used to infect U2OS cell line and selected stable TENT4A knockdown U2OS cells. Control shRNA infected U2OS cells were applying as control. To further knockdown, both stable TENT4A knockdown and control U2OS cells were transfected with SMARTpool siGENOME TENT4A siRNA (triplicates) and siGENOME Non-Targeting Control siRNA #5 (triplicates) using Lipofectamine RNAiMAX as recommended transfection protocols by the manufacturer. XP12RO cells were transfected with SMARTpool siGENOME TENT4A siRNA (triplicates) and siGENOME Non-Targeting Control siRNA #5 (triplicates) using Lipofectamine RNAiMAX as recommended transfection protocols by the manufacturer. After 48 h post-transfection, total RNA was extracted and expression profiles were analyzed in both U2OS and XP12RO.
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| Contributor(s) |
Swain U, Friedlander G, Livneh Z |
| Citation(s) |
34203408 |
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| Submission date |
Dec 11, 2019 |
| Last update date |
Jul 07, 2021 |
| Contact name |
Gilgi Friedlander |
| Organization name |
Weizmann Institute of Science
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| Street address |
Hertzel st
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| City |
Rehovot |
| ZIP/Postal code |
76100 |
| Country |
Israel |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA594987 |
| SRA |
SRP237261 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE141870_DEoutput_U2OS.xlsx |
2.1 Mb |
(ftp)(http) |
XLSX |
| GSE141870_DEoutput_XP_A.xlsx |
2.0 Mb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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