Abstract
During the recent years lysine methyltransferase Set7/9 ((Su(var)-3–9, Enhancer-of-Zeste, Trithorax) domain containing protein 7/9) has emerged as an important regulator of different transcription factors. In this study, we report a novel function for Set7/9 as a critical co-activator of E2 promoter-binding factor 1 (E2F1)-dependent transcription in response to DNA damage. By means of various biochemical, cell biology, and bioinformatics approaches, we uncovered that cell-cycle progression through the G1/S checkpoint of tumour cells upon DNA damage is defined by the threshold of expression of both E2F1 and Set7/9. The latter affects the activity of E2F1 by indirectly modulating histone modifications in the promoters of E2F1-dependent genes. Moreover, Set7/9 differentially affects E2F1 transcription targets: it promotes cell proliferation via expression of the CCNE1 gene and represses apoptosis by inhibiting the TP73 gene. Our biochemical screening of the panel of lung tumour cell lines suggests that these two factors are critically important for transcriptional upregulation of the CCNE1 gene product and hence successful progression through cell cycle. These findings identify Set7/9 as a potential biomarker in tumour cells with overexpressed E2F1 activity.
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Abbreviations
- DDR:
-
DNA damage response
- Set7/9:
-
(Su(var)-3–9, Enhancer-of-Zeste, Trithorax) domain containing protein 7/9
- E2F1:
-
E2 promoter-binding factor 1
- KMTase:
-
lysine methyltransferase
- TAF10:
-
TATA box binding protein (TBP)-associated factor, 30 kDa
- ERα:
-
oestrogen receptor
- PCAF:
-
P300/CBP-associated factor
- Rb:
-
retinoblastoma protein
- Cdk:
-
cyclin-dependent kinase
- BrdU:
-
bromodeoxyuridine
- siRNA:
-
small interfering RNA
- shRNA:
-
short hairpin RNA
- ChIP:
-
chromatin immunoprecipitation
- HDAC:
-
histone deacetylase
- HSP90:
-
heat shock protein 90
- PARP:
-
poly (ADP-ribose) polymerase
- RPMI:
-
Roswell Park Memorial Institute
- FBS:
-
foetal bovine serum
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- RIPA:
-
radioimmunoprecipitation assay
- SDS-PAGE:
-
SDS-polyacrylamide gel electrophoresis
- PBS:
-
phosphate buffer saline
- BSA:
-
bovine serum albumin
- FACS:
-
fluorescence-activated cell sorting
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Acknowledgements
This work was supported by MRC and by the Russian Ministry of Education (11.G34.31.0069) to GM and the Russian Foundation for Basic Research (A_2013 13-04-01024) and the Programme in Molecular and Cellular Biology from the Russian Academy of Sciences to NB.
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Lezina, L., Aksenova, V., Ivanova, T. et al. KMTase Set7/9 is a critical regulator of E2F1 activity upon genotoxic stress. Cell Death Differ 21, 1889–1899 (2014). https://doi.org/10.1038/cdd.2014.108
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DOI: https://doi.org/10.1038/cdd.2014.108
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