Diseases of the Reproductive System
Smale, Wendy1, Harris, Sarah2, Gersak, Ksenija3, Vincent, Andrea1, Shelling, Andrew1, 1 University of Auckand, Auckland, New Zealand2 University of Edinburgh, Edinburgh, Scotland3 University Medical Centre Ljubljana, Ljubljana, Slovenia
ABSTRACT- Premature ovarian failure (POF) is the occurrence of menopause before the age of 40 and affects approximately 1% of women worldwide. The majority of POF cases are idiopathic however some clearly have a genetic basis. The forkhead genes are an ever expanding family of transcription factors which are widely expressed in a range of tissues, have diverse roles in development and metabolism and have been implicated in various forms of cancer and disease. They contain a highly conserved 100 amino acid DNA binding (forkhead) domain. FOXL2 is a single exon gene expressed in the ovary and developing eyelid. Numerous FOXL2 mutations have been characterised in patients with BPES (blepharophimosis-ptosis-epicanthus inversus syndrome); a rare eyelid disorder which is often associated with POF. We have identified several novel FOXL2 mutations in our cohort of patients with isolated POF. Of particular interest are deletions in the polyalanine tract which may interfere with protein-protein interactions or alter protein secondary structure and hence affect its normal function. Constructs of wild type and variant forms of FOXL2 are being made and expressed in E.coli. DNA binding targets of FOXL2 will be identified by electromobility shift assays using putative target DNA binding sequences. These sequences will then be matched to the promoter regions of candidate target genes, leading to the identification of FOXL2-regulated pathways. The variant proteins will be tested against wild type FOXL2 to determine the effect of mutations on DNA binding and their potential role in POF. Determining the molecular basis of POF will lead to genetic tests and potential treatments.
KEY WORDS: premature ovarian failure, foxl2, forkhead transcription factor