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Experimental and bioinformatic analysis of cultured Bovine Endometrial Cells (BEND) responding to interferon tau (IFNT).

Palma-Vera SE, Einspanier R - Reprod. Biol. Endocrinol. (2016)

Bottom Line: Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6.All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.

ABSTRACT

Background: In ruminants, embryo implantation depends on progesterone (P4) and interferon tau (IFNT) controlling endometrial function. IFNT antagonizes bovine endometrial cells (BEND) response to phorbol 12,13-dibutyrate (PDBU) through posttranscriptional regulation of gene expression. We have previously described microRNAs (miRNAs) profiles in bovine endometrium, detecting miR-106a, relevant for embryo maternal communication. In this study, we investigated the expression miR-106a and genes for prostaglandin-endoperoxide synthase 2 (PTGS2), phospholipase A2, group IVA (PLA2G4A), estrogen receptor 1 (ESR1) and progesterone receptor (PR) in response to IFNT in BEND cells and searched for interferon responsive factors (IRFs) binding sites in their promoter genomic regions. The aim of this study was to unravel the molecular mechanisms involved in IFNT signalling and its regulation of miR-106a.

Findings: PTGS2 showed increased expression under PDBU, which was antagonized by IFNT. IFNT induced expression of PR and miR-106a and downregulation of ESR1 and PR. Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6. All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.

Conclusions: We report the IFNT regulatory effect on miR-106a expression through IRF-6 in bovine endometrial cells. We identified a set of potential binding sites for IRF-1 and IRF-6 within the bovine genome. A set of candidate gene regions could be characterized where IFNT can act via IRFs to regulate the expression of proteins and miRNAs. Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

No MeSH data available.


Related in: MedlinePlus

Sequence logos for IRF-1, IRF-2, IRF-6 and IRF-9 binding motifs. Sequences were retrieved from MotifDb and searched in the bovine genomic regions corresponding to the promoters of PTGS2, PLA2G4A, ESR1, PR and MIR106A
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Fig3: Sequence logos for IRF-1, IRF-2, IRF-6 and IRF-9 binding motifs. Sequences were retrieved from MotifDb and searched in the bovine genomic regions corresponding to the promoters of PTGS2, PLA2G4A, ESR1, PR and MIR106A

Mentions: We searched for the binding sites of IRFs in the bovine genome at the promoter site of known genes. Binding sites were determined by the presence of DNA motifs for a specific IRF. These motifs can be visualized as sequence logos in Fig. 3, showing the frequency of nucleotides at each position of the sequence. IRFs binding sites lengths ranged from 7 (IRF-6) to 18 (IRF-2), all having adenines as the most prevalent nucleotides. IRFs were selected based on previous studies, as they are known to be present in the endometrium of ruminants [5]. For protein coding genes, there were severe differences in the number of binding sites: IRF-6 was identified more than 40 thousand times, while IRF-1, 2 and 9 lay far behind (Table 3). A similar pattern was detected for miRNA coding genes. We decided to search for promoter binding sites at genes relevant for BEND function and miR-106a, leaving out thousands of genomic regions where IRFs can bind. These regions may regulate the expression of other genes and miRNAs. Future experimental studies will define what their roles are in order to detect pathways controlled by IFNT in BEND cells.Fig. 3


Experimental and bioinformatic analysis of cultured Bovine Endometrial Cells (BEND) responding to interferon tau (IFNT).

Palma-Vera SE, Einspanier R - Reprod. Biol. Endocrinol. (2016)

Sequence logos for IRF-1, IRF-2, IRF-6 and IRF-9 binding motifs. Sequences were retrieved from MotifDb and searched in the bovine genomic regions corresponding to the promoters of PTGS2, PLA2G4A, ESR1, PR and MIR106A
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835850&req=5

Fig3: Sequence logos for IRF-1, IRF-2, IRF-6 and IRF-9 binding motifs. Sequences were retrieved from MotifDb and searched in the bovine genomic regions corresponding to the promoters of PTGS2, PLA2G4A, ESR1, PR and MIR106A
Mentions: We searched for the binding sites of IRFs in the bovine genome at the promoter site of known genes. Binding sites were determined by the presence of DNA motifs for a specific IRF. These motifs can be visualized as sequence logos in Fig. 3, showing the frequency of nucleotides at each position of the sequence. IRFs binding sites lengths ranged from 7 (IRF-6) to 18 (IRF-2), all having adenines as the most prevalent nucleotides. IRFs were selected based on previous studies, as they are known to be present in the endometrium of ruminants [5]. For protein coding genes, there were severe differences in the number of binding sites: IRF-6 was identified more than 40 thousand times, while IRF-1, 2 and 9 lay far behind (Table 3). A similar pattern was detected for miRNA coding genes. We decided to search for promoter binding sites at genes relevant for BEND function and miR-106a, leaving out thousands of genomic regions where IRFs can bind. These regions may regulate the expression of other genes and miRNAs. Future experimental studies will define what their roles are in order to detect pathways controlled by IFNT in BEND cells.Fig. 3

Bottom Line: Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6.All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

View Article: PubMed Central - PubMed

Affiliation: Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.

ABSTRACT

Background: In ruminants, embryo implantation depends on progesterone (P4) and interferon tau (IFNT) controlling endometrial function. IFNT antagonizes bovine endometrial cells (BEND) response to phorbol 12,13-dibutyrate (PDBU) through posttranscriptional regulation of gene expression. We have previously described microRNAs (miRNAs) profiles in bovine endometrium, detecting miR-106a, relevant for embryo maternal communication. In this study, we investigated the expression miR-106a and genes for prostaglandin-endoperoxide synthase 2 (PTGS2), phospholipase A2, group IVA (PLA2G4A), estrogen receptor 1 (ESR1) and progesterone receptor (PR) in response to IFNT in BEND cells and searched for interferon responsive factors (IRFs) binding sites in their promoter genomic regions. The aim of this study was to unravel the molecular mechanisms involved in IFNT signalling and its regulation of miR-106a.

Findings: PTGS2 showed increased expression under PDBU, which was antagonized by IFNT. IFNT induced expression of PR and miR-106a and downregulation of ESR1 and PR. Bioinformatic analyses detected that PLA2G4A was associated to IRF-1 and IRF-6, while ESR1, PR and PTGS2 were associated to only IRF-6. All genes exhibit one motif per IRF, except miR-106a that had three binding sites for IRF-6.

Conclusions: We report the IFNT regulatory effect on miR-106a expression through IRF-6 in bovine endometrial cells. We identified a set of potential binding sites for IRF-1 and IRF-6 within the bovine genome. A set of candidate gene regions could be characterized where IFNT can act via IRFs to regulate the expression of proteins and miRNAs. Future studies will use these data to detect new IFNT regulatory mechanisms in the endometrium.

No MeSH data available.


Related in: MedlinePlus