转录因子PAX4调控湖羊FSHR基因转录活性

doi:10.11843/j.issn.0366-6964.2018.05.006

Abstract

Abstract:

The present study aimed to understand the sequence characterization and transcriptional regulation of the 5'-UTR of Hu sheep FSHR gene, thus provided the basis for further exploring of transcription regulatory mechanism of FSHR gene. Adult female Hu sheep (n=3) were slaughtered for 11 tissues sampling of heart, liver, spleen, lung, kidney, stomach, muscle, rectum, intestinal, uterus and ovary. PCR amplification and clone sequencing were performed to determine the 5'-UTR sequence, and then the sequence characterization of Hu sheep FSHR gene was analyzed by bioinformatic method. Tissue expression patterns of transcription factor PAX4 of Hu sheep were detected by RT-PCR. Overexpression vector (pcDNA3.1-PAX4) of Hu sheep PAX4 gene was synthesized, which was then transfected into pig ovarian granulosa cells (GCs), and the apoptosis rate of GCs was measured by flow cytometry. The dul-luciferase reporter assay system was used to evaluate the effect of PAX4 on transcriptional activity of Hu sheep FSHR gene. The results showed that the full length of the 5'-UTR of Hu sheep FSHR gene was 161 bp, which contained several typical transcriptional regulatory elements such as E-box, CAAT-box and GC-box. A few of binding sites for transcription factor were also found, such as GATA-2, Sp1 and PAX4. RT-PCR showed that transcription factor PAX4 was expressed in 11 tissues of Hu sheep, and moderately expressed in ovarian tissue. After transfecting with pcDNA3.1-PAX4, the mRNA levels of PAX4 was increased (P<0.01) in GCs. Overexpression of PAX4 significantly increased the apoptosis rate of GCs (P<0.05), and decreased the luciferase activity of luciferase reporter vectors containing FSHR 5'-UTR in both GCs (P<0.05) and COS-7 cells (P<0.01). Together, the findings demonstrated that transcription factor PAX4 could enhance GC apoptosis by inhibiting transcriptional activity of FSHR gene in Hu sheep.

CLC Number:

S826.2

WANG De-di, LI Yin-xia, YAO Yi-long, PAN Zeng-xiang, JUE Ken, YI Ming, CAO Shao-xian, LI Qi-fa. Transcription Factor PAX4 Regulates the Transcriptional Activity of FSHR Gene in Hu Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 919-926.

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Transcription Factor PAX4 Regulates the Transcriptional Activity of FSHR Gene in Hu Sheep

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