Mutagenesis of RRL-NSD3-Short-3xFLAG Lentiviral Vector

SGC Open Notebook Project to Characterize the HMTase NSD3

Exp013 Objective: The NSD3’s PWWP1 domain is present in both long and short isoforms. It has been shown to bind H3K36me2 (Sankaran et al.(2016) - PMID:26912663) and be required for the maintenance of AML (Chen et al.(2015 - PMID: 26912663). However, it is still unclear how this domain contributes to NSD3’s function at enhancers. To study this aspect of NSD3’s biology, I will mutate W284 to alanine in the RRLNSD3-3xFLAG-IRES-Puro plasmids I described earlier (exp010) by site-directed mutagenesis. This is the second tryptophan within the PWWP1 motif and is critical for substrate recognition (Qin, S & Min, J(2014) - PMID:25277115). This construct will be useful for understanding how NSD3 recognition of methylated histones influences its putative activity in cancer.