Determinants Of Faim Transcriptional Regulation
INTRODUCTION: Recent work by the WMed Center for Immunobiology demonstrated the ability of Fas Apoptosis Inhibitory Molecule (FAIM) to combat cellular stress.
OBJECTIVE: We sought to explore FAIM's transcriptional regulation for insight into when, where, and how FAIM is produced.
METHODS: Firefly Luciferase transcription reporters of varying lengths were created and tested in HEK 293T cell lines subjected to heat stress. Differences in reporter expression were monitored, and allowed identification of potential FAIM regulatory regions.
Transcription factor candidates for the regions of interest were identified via two methods. First, UCSC's Genome Browser database provided candidates that show any binding to the FAIM gene, and potential binding sites were identified by comparing FAIM base pairs to the transcription factors' consensus binding sequences. Second, we used MEME-SUITE prediction software to identify additional transcription factor candidates.
RESULTS: Two regions on the FAIM gene, -250→+75 and +900→+1100, were determined to regulate transcription. The former acts as a promoter, especially after heat stress, and the latter works as a repressor. Genome Browser and MEME-SUITE allowed us to identify transcription factors that potentially bind these regions: JUND, RUNX3, and TEAD4.
DISCUSSION: This work points the way to elucidating FAIM regulation. The regions of interest will be subjected to chromatin immunoprecipitation assays to definitively assess the role of candidate transcription factors.
In this work, we have identified two regions of interest and corresponding candidate transcription factors that may be involved in regulating this novel gene that plays a role in the cellular stress responses.