A Study On The Mechanism Of The Nuclear Transportation Of Neurogenic Differentiation Factor NeuroD2 Of The BHLH Families

NeuroD2,a transcription factor in the neurogenic differentiation Neuro D Protein subfamily,is a member of the bHLH differentiation factors.It can bind with E-BOX and activate the transcription of its target genes.Neuro D 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,such as DNAMAN.This experiment intend to draw a conclusion that gene and protein sequences of NeuroD2 in the Neuro D family of human beings have the highest homology with those of mice.Thus an assumption can be concluded that NeuroD2 may have a great value to the development of nerve cells.In the experiment,This experiment intend to firstly determined the subcellular localization of endogenous NeuroD2 protein in mice.And then This experiment intend to have proved that 14-3-3epsilonepsilonis an interacting protein of NeuroD2 via pull-down assay.To investigate whether 14-3-3epsilonepsilon protein will affect the subcellular localization of NeuroD2,eukaryotic expression vector of NeuroD2 and 14-3-3epsilonepsilon-RNAi expression vector were constructed.And then these vectors transfected separately or co-transfected Hela cells.At last Western blot was performed to analys the distribution of NeuroD2 protein.Finally the result show that after 14-3-3epsilonepsilon being suppressed,the efficiency of nuclear transportation of NeuroD2 has decreased a lot in the absent of 14-3-3epsilonepsilon protein,but it is not totally inhibited.According to the analysis,the possible reasons may be the influence of residual 14-3-3epsilonepsilon protein in cells or the interaction of other proteins which have the similar function of nuclear transportation.