Senataxin Plays an Essential Role with DNA Damage Response Proteins in Meiotic Recombination and Gene Silencing

Abstract

Senataxin, mutated in the human genetic disorder ataxia with oculomotor apraxia type 2 (AOA2), plays an important role in maintaining genome integrity by coordination of transcription, DNA replication, and the DNA damage response. We demonstrate that senataxin is essential for spermatogenesis and that it functions at two stages in meiosis during crossing-over in homologous recombination and in meiotic sex chromosome inactivation (MSCI). Disruption of the Setx gene caused persistence of DNA double-strand breaks, a defect in disassembly of Rad51 filaments, accumulation of DNA:RNA hybrids (R-loops), and ultimately a failure of crossing-over. Senataxin localised to the XY body in a Brca1-dependent manner, and in its absence there was incomplete localisation of DNA damage response proteins to the XY chromosomes and ATR was retained on the axial elements of these chromosomes, failing to diffuse out into chromatin. Furthermore persistence of RNA polymerase II activity, altered ubH2A distribution, and abnormal XY-linked gene expression in Setx ^−/− revealed an essential role for senataxin in MSCI. These data support key roles for senataxin in coordinating meiotic crossing-over with transcription and in gene silencing to protect the integrity of the genome.

Author Summary

Ataxia with oculomotor apraxia type 2 (AOA2) caused by a defect in the gene Setx (coding for senataxin) is part of a subgroup of autosomal recessive ataxias characterized by defects in genes responsible for the recognition and/or repair of damage in DNA. Cells from these patients are characterized by oxidative stress and are defective in RNA processing and termination of transcription. Recent data suggest that senataxin is involved in coordinating events between DNA replication forks and ongoing transcription. To further understand the role of senataxin, we disrupted the Setx gene in mice and demonstrated its essential role in spermatogenesis during meiotic recombination and in meiotic sex chromosome inactivation (MSCI). In the absence of senataxin, DNA double-strand breaks persist, RNA:DNA hybrids (R-loops) accumulate, and homologous recombination is disrupted. Senataxin localised to the XY chromosomes during pachytene. This was dependent on Brca1, which functions early in MSCI to recruit DNA damage response proteins to the XY body. In the absence of senataxin, there was incomplete accumulation of DNA damage response proteins on the XY chromosomes and no MDC1-dependent diffusion of ATR to the broader XY chromatin. The end result was a defect in MSCI, apoptosis, and a failure to complete meiosis.