Neurodegeneration and aging: roles for DNA single-strand break repair. (360G-Wellcome-085284_Z_08_Z)

The SSBR proteins TDP1 and Aprataxin are mutated in two distinct neurological diseases; spinocerebellar ataxia with axonal neuropathy-1 (SCAN1) and ataxia oculomotor apraxia-1 (AOA1). The goal of this proposal is to (1) Characterise mechanisms of SSBR in primary neural cells using a combination of in vitro biochemical assays (employing synthetic oligonucleotide substrates) and cellular chromosomal DNA repair assays (employing the alkaline comet assays). I propose to define the intermediates that accumulate in neural cells that are defective in SSBR using biochemical approaches and to characterise novel factors in this pathway using yeast 2-hybrid and co-immunoprecipitation experiments. (2) Elucidate the impact of un-repaired SSBs on neural cell fate, with emphasis on transcriptional competence and survival of primary neural cells. (3) Evaluate the neuroprotective effect of SSBR in vivo, by measuring specific behavioural and physiological end points in wild type and SSBR-defective mouse model systems.