Characteristics of the BMP7 Promoter in Hu Sheep.

Xiaoyang Lv,Ling Chen,Shuangxia Zou,Wei Sun,Jinyu Wang,Joram M Mwacharo

doi:10.3390/ani9110874

Abstract

Simple SummaryBone morphogenetic protein 7 (BMP7) is one of the largest secretory signal conductive molecules and is in the TGF-β superfamily. It plays an important role in the growth and development of hair follicles. We cloned the proximal promoter of the BMP7 gene for bioinformatics analysis. Dual-luciferase reporter system and overexpression were used to analyze the key regions and transcription factor binding sites. There was high activity between −758 bp and −545 bp in the core region of the gene and a possible binding site for transcription factors SP1 and EGR1.The BMP7 gene is involved in the growth and development of hair follicles but its regulation mechanism is unclear. We studied the regulation mechanism of the BMP7 promoter by cloning the proximal promoter of BMP7 for bioinformatics analysis. A series of missing vectors was then constructed for dual-fluorescein activity detection based on the bioinformatics analysis results. We tested transcription-factor binding-site mutations and transcription factor over-expression to analyze the transcriptional regulation principle of the BMP7 promoter region. The upstream transcriptional regulatory region of the BMP7 gene proximal promoter was predicted by bioinformatics. There were −1216 bp to −1166 bp and −632 bp to −582 bp transcription initiation sites in the upstream transcriptional regulatory region of the BMP7 gene proximal promoter. The CpG islands’ distribution showed that there were many CpG islands at −549 bp to 1 bp. A dual-luciferase assay revealed high activity between −758 bp and −545 bp in the core region and a possible binding site for transcription factors SP1 and EGR1. The transcriptional activity of BMP7 was significantly decreased in the transcriptional regulatory region of the BMP7 after EGR1 and SP1 mutation. Transcription was significantly enhanced by over expression of the EGR1 transcription factor, which strongly suggests that EGR1 and SP1 play important roles in BMP7 regulation.

Highlights

Bone morphogenetic protein (BMP) belongs to the transforming growth factor-β (TGF-β) superfamily [1]
In noggin-knockout mice, prolonged periods of excessive BMPs impeded the development of primary hair follicles and affected the formation process of inner root sheaths and hair shafts [5,6]
Wang [9] noted that BMP7 can inhibit wool growth in fine wool sheep. These results indicate that BMP7 may promote the proliferation of hair follicle cells in the hair follicle growth phase

Summary

Introduction

Bone morphogenetic protein (BMP) belongs to the transforming growth factor-β (TGF-β) superfamily [1]. The BMP family has many biological functions in cell proliferation, differentiation, bone formation, tumors, and hair follicle growth. The BMP signaling pathway may inhibit the Wnt/β-catenin signaling pathways to promote hair cycle growth [2] and enable the hair follicle stem cells in the Animals 2019, 9, 874; doi:10.3390/ani9110874 www.mdpi.com/journal/animals. The BMP signaling pathway is critical for the differentiation of hair root sheaths and hair shafts. Kulessa [4] found that transgenic mice overexpressed the BMP antagonist noggin caused hair matrix cells proliferation and hair precursor cells differentiation leading to hair growth. In noggin-knockout mice, prolonged periods of excessive BMPs impeded the development of primary hair follicles and affected the formation process of inner root sheaths and hair shafts [5,6]

Methods

Results

Discussion

Conclusion