766 / 2019-06-14 15:12:43

Using the nuclear transcriptional regulator RCD1, to find PARylated & PAR-related proteins.

PAR,RCD1,transcriptional regulation,poly-ADP-ribose,PAR-binding

全体主题 > Signal Transduction

The posttranslational modification poly-ADP-ribose (PAR) is a transient modification of proteins, which is linked with chromatin remodeling programmed cell death amongst others. This transient chain of APD-ribose monomers is synthesized by poly-ADP-ribose polymerases. Recently, some mysteries about PARylated proteins have been solved in the animal field. In plants, however, many mechanisms about PARylation and PARylated proteins and the functioning thereof have remained in the dark. This work aims to characterize in vivo proteins that have undergone addition of PAR and study the functional aspect of this event. Extraction and enrichment of PAR are performed using the two domains responsible for PAR-binding of RCD1, the WWE- and PARP-like domain. RADICAL CELL DEATH 1 (RCD1) is a plant protein that has a strong affinity to PAR in vitro and possesses another C-terminal domain that allows it to interact with over 30 transcription factors. As such, RCD1 has the potential to be a PAR reader, which upon PAR-binding might change its interaction with these transcription factors. The corresponding domains of the closest homolog of RCD1, SIMILAR TO RCD ONE 1 (SRO1), will also be employed for enrichment. After which mass spectrometry will be performed to identify the enriched proteins, amongst which will be PARylated proteins and PAR associated proteins. Which opens the door to study how this modification affects its targets.