Manipulating Cellular Trafficking Positively Affects Syn‐TEF Function in Human Tissue

Abstract

Synthetic transcription elongation factors (Syn‐TEFs) bind to the minor groove of DNA and recruit transcription machinery. Syn‐TEFs can provide therapeutic relief for transcriptionally defective disorders such as Friedreich's ataxia, a neurodegenerative disease resulting from a 90% loss in Frataxin (FXN) expression. Syn‐TEF1 restores FXN expression in Friedreich's ataxia patient cells. However, the mechanism of cellular entry has yet to be characterized. From our observations, only a fraction of the compound is available to act in the nucleus. Fluorescently tagged Syn‐TEF analogues aggregate into bright puncta in the cytoplasm of imaged cells. Using confocal microscopy and reverse transcription quantitative polymerase chain reaction (RT‐qPCR), we assessed conditions of puncta formation and tested means of resolving or preventing the aggregation of Syn‐TEFs into puncta in human cells. We have found that manipulating cellular uptake and trafficking can improve nuclear localization of fluorescent Syn‐TEF analogues and change Syn‐TEF1 effective concentrations. Understanding the mechanisms of uptake for Syn‐TEFs and their fluorescent analogues will provide insight into how Syn‐TEFs affect gene expression in Friedreich's ataxia and related disorders.Support or Funding InformationThis project was supported by NIH NINDS grant R01NS108376 and the Clinical and Translational Science Award (CTSA) program through the National Center for Advancing Translational Sciences (NCATS), grant UL1TR002373. S.R. was also supported by the Trewartha Senior Thesis Research Grant.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.