Changes in splicing factor expression are associated with advancing age in man

Abstract

Human ageing is associated with decreased cellular plasticity and adaptability. Changes in alternative splicing with advancing age have been reported in man, which may arise from age-related alterations in splicing factor expression. We determined whether the mRNA expression of key splicing factors differed with age, by microarray analysis in blood from two human populations and by qRT-PCR in senescent primary fibroblasts and endothelial cells. Potential regulators of splicing factor expression were investigated by siRNA analysis. Approximately one third of splicing factors demonstrated age-related transcript expression changes in two human populations. Ataxia Telangiectasia Mutated (ATM) transcript expression correlated with splicing factor expression in human microarray data. Senescent primary fibroblasts and endothelial cells also demonstrated alterations in splicing factor expression, and changes in alternative splicing. Targeted knockdown of the ATM gene in primary fibroblasts resulted in up-regulation of some age-responsive splicing factor transcripts. We conclude that isoform ratios and splicing factor expression alters with age in vivo and in vitro, and that ATM may have an inhibitory role on the expression of some splicing factors. These findings suggest for the first time that ATM, a core element in the DNA damage response, is a key regulator of the splicing machinery in man.

Keywords: ATM; Ageing; Ataxia Telangiectasia Mutated gene; Heterogeneous nuclear ribonucleoproteins; Human; LMNA; Lamin A/C gene; RT-PCR; Reverse Transcription PCR; SR splicing factor; SRSF; Splicing; TLDA; TaqMan Low Density Array; hnRNP; qRT-PCR; quantitative real-time PCR; siRNA; small interfering RNA.