Transcriptional Activity of the FUT1 Gene Promoter Region in Pigs

Abstract

This study aims to provide a theoretical basis on the regulatory mechanism of the α-l,2-fucosyltransferase (FUT1) gene in pigs by analyzing the transcriptional activity of its promoter region. On the basis of the previously obtained promoter sequence, primers upstream and downstream of the gene were designed using the restriction endonucleases KpnI and HindIII respectively, and the recombinant plasmids of the pGL3-promoter were constructed by inserting promoter sequences with partially missing regions. The resultant mutants were observed by transient transfection assay into HEK293 cells, and the transcriptional activity of the promoter region was determined by luciferase activity. The 5′-flanking region of the FUT1 gene (−1150 to +50 bp) exhibited promoter activity. The −1150-bp to −849-bp region showed negative regulation of the gene. The recombinant plasmid pGL3-898 showed the strongest luciferase activity, and the activity showed a decreasing trend when the deleted region was increased. Recombinant plasmids were successfully constructed, verified, and the positive and negative regulation areas and core promoter region were detected, providing a deeper insight into the transcriptional regulatory mechanism of the FUT1 gene.

Hypoxia inducible factor 1α down regulates cell surface expression of α1,2‐fucosylated glycans in human pancreatic adenocarcinoma cells

“…In addition, HIF-1a expression was inversely correlated with FUT1/2 expression and cell surface fucosylation in PaTu-S cells, and in the colon cancer cell lines SW480 and HTC116, indicating that this effect is not restricted to PDAC cells. Our data are not directly in accordance with recent data which show that FUT1 transcription was elevated under hypoxic conditions in prostate cancer cells [31]. Whereas hypoxia is known to induce HIF-1a expression, the role of HIF-1a in the regulation of FUT1 was not investigated in the latter study, leaving the possibility open that other hypoxia-induced factors could be involved in the observed upregulation of FUT1 in prostate cancer [32].…”

“…The mechanism by which HIF‐1α suppresses FUT1/2 expression has not been clarified in this work, and future studies are required to fully understand the function of this transcription factor in the suppression of α1,2‐fucosylation, and its relevance for cell migration and metastasis. Interestingly, results of studies on the transcriptional regulation of the pig FUT1 gene demonstrated that this gene possesses positive and negative regulatory elements in the upstream non‐coding region [31]. The negative regulatory area impaired transcription initiation even though abundant cis ‐acting elements and transcription factor‐binding loci were predicted in that specific area of the FUT1 promoter gene [31].…”

“…Interestingly, results of studies on the transcriptional regulation of the pig FUT1 gene demonstrated that this gene possesses positive and negative regulatory elements in the upstream non‐coding region [31]. The negative regulatory area impaired transcription initiation even though abundant cis ‐acting elements and transcription factor‐binding loci were predicted in that specific area of the FUT1 promoter gene [31]. Since the human FUT1 and FUT2 contain multiple HIF transcription factor consensus sequences (data not shown) it is possible that HIF‐1α binds directly to regulatory repressor regions in the promoter of these genes, thereby preventing their transcription.…”

Promoter identification and analysis of key glycosphingolipid biosynthesis-globo series pathway genes in piglets

“…IPEC-J2 cells were transfected with recombinant plasmids pGL3-P1 and pGL3-P2 for 24 h (Zi et al, 2013), and dual luciferase assays were performed. The products were subjected to electrophoresis on a 1% agarose gel.…”

Detection of functional polymorphisms influencing the promoter activity of the SAA2 gene and their association with milk production traits in Chinese Holstein cows

“…Along with the rapid development and reduced costs of next-generation sequencing (NGS), high-throughput RNA sequencing (RNA-seq) has rapidly emerged as a major transcriptome profiling system to discover differentially expressed genes and novel transcripts (Garber et al 2011). So far, there have been some RNA-seq studies on human (Zi et al 2013), chicken (Katie et al 2013) and pig (Chen et al 2011). Also, RNAseq has been successfully employed in bovine embryo (Huang & Khatib 2010) and milk transcriptome investigations (Canovas et al 2010).…”