Preliminary Study On The Transcriptional Regulation Of MicroRNA PC-3P-2869 By Transcription Factor ELK1

Deer antler,as a special epiphyte with periodic growth characteristics,has high biomedical value.Heritable alterations in gene expression,without any alteration in genes,are referred to as epigenetics.MicroRNA(miRNA)is a key regulator of epigenetics.In our previous study,a novel miRNA,named miR-PC-3P-2869(miR-PC-2869),was identified during the rapid growth phase of red deer(Cervus elaphus),but the regulatory mechanism of its transcription and expression remains unclear.The location of miR-PC-2869 in the reference genome of red deer was mapped,and the core promoters and transcription factors in its upstream regulatory region were predicted by bioinformatics analysis.To confirm the results,we performed luciferase reporter gene tests.The results are shown below:(1)The CDS region of ELK1 protein of northeastern red deer was obtained for the first time.Bioinformatics analysis showed that the entire CDS region of ELK1 was 1323 bp,which had strong evolutionary conservation and had the highest similarity with bovine ELK1 gene(97.99%).The CDS region of the ELK1 genome encoded a 440-amino acid ELK1 protein with a molecular weight of 46.2 KD.ELK1 protein is hydrophilic with an isoelectric point of 5.81.Its secondary structure includes random coil,α-helix,extended strand and β-fold.It has ETS domain and is located in the nucleus;(2)The core promoter region F is the core promoter region of miR-PC-2869;(3)The transcription factor ELK1 positively regulated the expression of miR-PC-2869.There were three binding sites in the promoter region of miR-PC-2869,and the three binding sites played a dominant role.This finding provides important basic data for the regeneration of deer antler cartilage tissue in northeastern red deer,which can be used as a basis for subsequent studies.