A Study On The Mechanism Of The Nuclear Transportation Of Neurogenic Differentiation Factor Neurod6

NeuroD6,a transcription factor in the neurogenic differentiation NeuroD Protein subfamily,is a member of the bHLH differentiation factors.It can bind with E-BOX and activate the transcription of its target genes.NeuroD family plays an important role in the process of regulating the embryonic brain tissue's development and in the survival and regeneration of nerve cells of adult animals.However,its specific regulation mechanism has not been discovered.Through the comparative analysis by using bioinformatics method together with related biological software,we draw a conclusion that gene and protein sequences of NeuroD6 in the NeuroD family of human beings has the highest homology with those of mice.Thus an assumption can be concluded that NeuroD6 may play a more important role in the development of nerve cells.In the experiment,we firstly determined the subcellular localization of endogenous NeuroD6 protein in mice.And then we have proved that ATW is an interacting protein of NeuroD6 via pull-down assay.To investigate whether ATW protein will affect the subcellular localization of NeuroD6,eukaryotic expression vector of NeuroD6 and ATW-RNAi expression vector were constructed.And then these vectors transfected separately or co-transfected Hela cells.At last Western blot was performed to analysis the distribution of NeuroD6 proteins.Finally the results show that after ATW being suppressed,the efficiency of nuclear transportation of NeuroD6 has decreased a lot in the absent of ATW protein,but it is not totally inhibited.According to the analysis,the possible reasons may be the influence of residual ATW protein in cells or the interaction of other proteins which have the similar function of nuclear transportation.