ELF3 activated by a superenhancer and an autoregulatory feedback loop is required for high-level HLA-C expression on extravillous trophoblasts
| Autor: | Sarika Kshirsagar, Sai Ma, Qin Li, Jack L. Strominger, Jason D. Buenrostro, Fang Wang, Tamara Tilburgs, Torsten B. Meissner, Ziming Du, Motonari Uesugi |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Small interfering RNA Indoles HLA-C superenhancer Adamantane MED1 Mediator Complex Subunit 1 0302 clinical medicine Immunology and Inflammation autoregulatory feedback loop Pregnancy RNA Small Interfering Promoter Regions Genetic Feedback Physiological Multidisciplinary Mediator Complex Chemistry Gene Expression Regulation Developmental Azepines Biological Sciences Cell biology Chromatin Trophoblasts DNA-Binding Proteins Enhancer Elements Genetic 030220 oncology & carcinogenesis embryonic structures Abortion Legal Female Protein Binding Signal Transduction Primary Cell Culture Regulatory Factor X Transcription Factors HLA-C Antigens Cell Line 03 medical and health sciences Coactivator Humans (+)-JQ1 Gene Transcription factor Proto-Oncogene Proteins c-ets Promoter Triazoles ELF3 Pregnancy Trimester First 030104 developmental biology HLA-B Antigens Chromatin immunoprecipitation Immunity Maternally-Acquired Transcription Factors |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America |
| ISSN: | 1091-6490 |
| Popis: | Significance Several techniques have identified ELF3 as particularly important in regulating trophoblast-specific HLA-C expression. This results from a single-nucleotide difference in the promotor of the HLA-C gene at the RFX5 binding site that creates an ELF3 binding site not found in the HLA-A or HLA-B promoters. We discovered a superenhancer and a positive autoregulatory feedback loop that promotes expression of the ELF3 gene in trophoblasts. Disruption of either the superenhancer by (+)-JQ1 or interference with the positive-feedback loop by wrenchnolol decreased ELF3 levels, and thus HLA-C expression. Aberrations of this complex regulatory system could be involved in control of infection, miscarriage, preterm birth, preeclampsia, as well as parturition in normal pregnancy and in development of choriocarcinoma. HLA-C arose during evolution of pregnancy in the great apes 10 to 15 million years ago. It has a dual function on placental extravillous trophoblasts (EVTs) as it contributes to both tolerance and immunity at the maternal–fetal interface. The mode of its regulation is of considerable interest in connection with the biology of pregnancy and pregnancy abnormalities. First-trimester primary EVTs in which HLA-C is highly expressed, as well as JEG3, an EVT model cell line, were employed. Single-cell RNA-seq data and quantitative PCR identified high expression of the transcription factor ELF3 in those cells. Chromatin immunoprecipitation (ChIP)-PCR confirmed that both ELF3 and MED1 bound to the proximal HLA-C promoter region. However, binding of RFX5 to this region was absent or severely reduced, and the adjacent HLA-B locus remained closed. Expression of HLA-C was inhibited by ELF3 small interfering RNAs (siRNAs) and by wrenchnolol treatment. Wrenchnolol is a cell-permeable synthetic organic molecule that mimics ELF3 and is relatively specific for binding to ELF3’s coactivator, MED23, as our data also showed in JEG3. Moreover, the ELF3 gene is regulated by a superenhancer that spans more than 5 Mb, identified by assay for transposase-accessible chromatin using sequencing (ATAC-seq), as well as by its sensitivity to (+)-JQ1 (inhibitor of BRD4). ELF3 bound to its own promoter, thus creating an autoregulatory feedback loop that establishes expression of ELF3 and HLA-C in trophoblasts. Wrenchnolol blocked binding of MED23 to ELF3, thus disrupting the positive-feedback loop that drives ELF3 expression, with down-regulation of HLA-C expression as a consequence. |
| Databáze: | OpenAIRE |
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