Expression of human frataxin is regulated by transcription factors SRF and TFAP2.
BACKGROUND
Friedreich ataxia is an autosomal recessive neurodegenerative disease caused by reduced expression levels of the frataxin gene (FXN) due to expansion of triplet nucleotide GAA repeats in the first intron of FXN. Augmentation of frataxin expression levels in affected Friedreich ataxia patient tissues might substantially slow disease progression.


RESULTS
We utilized bioinformatic tools in conjunction with chromatin immunoprecipitation and electrophoretic mobility shift assays to identify transcription factors that influence transcription of the FXN gene. We found that the transcription factors SRF and TFAP2 bind directly to FXN promoter sequences. SRF and TFAP2 binding sequences in the FXN promoter enhanced transcription from luciferase constructs, while mutagenesis of the predicted SRF or TFAP2 binding sites significantly decreased FXN promoter activity. Further analysis demonstrated that robust SRF- and TFAP2-mediated transcriptional activity was dependent on a regulatory element, located immediately downstream of the first FXN exon. Finally, over-expression of either SRF or TFAP2 significantly increased frataxin mRNA and protein levels in HEK293 cells, and frataxin mRNA levels were also elevated in SH-SY5Y cells and in Friedreich ataxia patient lymphoblasts transfected with SRF or TFAP2.


CONCLUSIONS
We identified two transcription factors, SRF and TFAP2, as well as an intronic element encompassing EGR3-like sequence, that work together to regulate expression of the FXN gene. By providing new mechanistic insights into the molecular factors influencing frataxin expression, our results should aid in the discovery of new therapeutic targets for the treatment of Friedreich ataxia.
Li
Kuanyu
K
Jiangsu Key Laboratory for Molecular Medicine, Medical School of Nanjing University, Nanjing, China. likuanyu@nju.edu.cn
Singh
Anamika
A
Crooks
Daniel R
DR
Dai
Xiaoman
X
Cong
Zhuangzhuang
Z
Pan
Liang
L
Ha
Dung
D
Rouault
Tracey A
TA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D019295:Computational Biology; D005621:Friedreich Ataxia / Q000473:pathology / Q000628:therapy; D005786:Gene Expression Regulation; D006801:Humans; D007501:Iron / Q000378:metabolism; D033862:Iron-Binding Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D026362:Serum Response Factor / Q000378:metabolism*; D050656:Transcription Factor AP-2 / Q000378:metabolism*; D015854:Up-Regulation; D000096923:Frataxin
D007501:Iron; D033862:Iron-Binding Proteins; D012333:RNA, Messenger; D026362:Serum Response Factor; D050656:Transcription Factor AP-2; D000096923:Frataxin; C476858:SRF protein, human
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