SIRT1 gene expression upon genotoxic damage is regulated by APE1 through nCaRE-promoter elements

Molecular Biology of the Cell
Giulia AntonialiGianluca Tell

Abstract

Apurinic/apyrimidinic endonuclease 1 (APE1) is a multifunctional protein contributing to genome stability via repair of DNA lesions via the base excision repair pathway. It also plays a role in gene expression regulation and RNA metabolism. Another, poorly characterized function is its ability to bind to negative calcium responsive elements (nCaRE) of some gene promoters. The presence of many functional nCaRE sequences regulating gene transcription can be envisioned, given their conservation within ALU repeats. To look for functional nCaRE sequences within the human genome, we performed bioinformatic analyses and identified 57 genes potentially regulated by APE1. We focused on sirtuin-1 (SIRT1) deacetylase due to its involvement in cell stress, including senescence, apoptosis, and tumorigenesis, and its role in the deacetylation of APE1 after genotoxic stress. The human SIRT1 promoter presents two nCaRE elements stably bound by APE1 through its N-terminus. We demonstrate that APE1 is part of a multiprotein complex including hOGG1, Ku70, and RNA Pol II, which is recruited on SIRT1 promoter to regulate SIRT1 gene functions during early response to oxidative stress. These findings provide new insights into the role of nCaRE sequen...Continue Reading

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Citations

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Methods Mentioned

BETA
phylogenetic footprinting
electrophoretic mobility shift assay
surface plasmon resonance
chip
transfection
footprinting
immunoprecipitation
reverse transcription-PCR
cleavage
PCR

Software Mentioned

Fasta3
GraphPad
mfold
Excel
Prism
BioMart
Perl
Glsearch
Ensembl
Evaluation

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