Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
5-Year Impact Factor – 2.2
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Index Copernicus  – 161.11; MEiN – 140 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2020, vol. 29, nr 6, June, p. 677–682

doi: 10.17219/acem/122168

Publication type: original article

Language: English

License: Creative Commons Attribution 3.0 Unported (CC BY 3.0)

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MicroRNA-548k upregulates a spliced variant of human CD2-associated protein by targeting its promoter

Nannan Li1,B,F, Guoping Zhou1,A

1 Department of Pediatrics, First Affiliated Hospital, Nanjing Medical University, China


Background. MicroRNAs (miRNAa) regulate gene expression by repressing the translation or directing the degradation of mRNA in the cytoplasm. However, recent studies have shown that miRNAs act to regulate transcription at target gene promoters.
Objectives. To investigate whether miR-548k regulates CD2AP002 expression at the transcriptional level.
Material and Methods. The CD2AP002 promoter region was cloned and its promoter activity was examined in HEK293 cells using luciferase assay. The CD2AP002 promoter region was characterized bioinformatically. The HEK293 cells were treated with a miR-548k mimic or inhibitor and the luciferase activity of the CD2AP002 promoter and the mRNA expression of CD2AP002 were assayed. A chromatin immunoprecipitation assay was performed with antibody against RNA polymerase II at the CD2AP002 promoter region in HEK293 cells.
Results. The CD2AP002 promoter region contained 2 putative miR-548k binding sites, and mutational analysis revealed that both of those sites contributed to transcriptional activity. The miR-548k mimics increased CD2AP002 promoter activity and mRNA expression. In contrast, the miR-548k inhibitor significantly reduced CD2AP002 promoter activity and mRNA expression. Finally, the chromatin immunoprecipitation assay showed that the enrichment of RNA polymerase II at the CD2AP002 promoter region was enhanced in the HEK293 cells treated with the miR-548k mimic.
Conclusion. This is the first study to demonstrate that miR-548k upregulates CD2AP002 expression by targeting its promoter.

Key words

transcriptional regulation, gene promoters, CD2-associated protein, microRNA-548k, spliced variant

References (22)

  1. Dragovic D, Rosenstock JL, Wahl SJ, Panagopoulos G, DeVita MV, Michelis MF. Increasing incidence of focal segmental glomerulosclerosis and an examination of demographic patterns. Clin Nephrol. 2005;63(1):1–7.
  2. Wolf G, Stahl RA. CD2-associated protein and glomerular disease. Lancet. 2003;362(9397):1746–1748.
  3. Shih NY, Li J, Karpitskii V, et al. Congenital nephrotic syndrome in mice lacking CD2-associated protein. Science. 1999;286(5438):312–315.
  4. Johnson JM, Castle J, Garrett-Engele P, et al. Genome-wide survey of human alternative pre-mRNA splicing with exon junction microarrays. Science. 2003;302(5653):2141–2144.
  5. Radmilovic M, Zukic B, Stankovic B, et al. Thalassemia syndromes in Serbia: An update. Hemoglobin. 2010;34(5):477–485.
  6. Andreadis A. Tau gene alternative splicing: Expression patterns, regulation and modulation of function in normal brain and neurodegenerative diseases. Biochim Biophys Acta. 2005;1739(2–3):91–103.
  7. Bartel DP. MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–297.
  8. Kalantari R, Chiang CM, Corey DR. Regulation of mammalian transcription and splicing by nuclear RNAi. Nucleic Acids Res. 2016;44(2):524–537.
  9. Paugh SW, Coss DR, Bao J, et al. MicroRNAs form triplexes with double stranded DNA at sequence-specific binding sites: An eukaryotic mechanism via which microRNAs could directly alter gene expression. PLoS Comput Biol. 2016;12(2):e1004744.
  10. Suzuki K, Ahlenstiel C, Marks K, Kelleher AD. Promoter targeting RNAs: Unexpected contributors to the control of HIV-1 transcription. Mol Ther Nucleic Acids. 2015;4(1):e222.
  11. Cho S, Park JS, Kang YK. AGO2 and SETDB1 cooperate in promoter-targeted transcriptional silencing of the androgen receptor gene. Nucleic Acids Res. 2014;42(22):13545–13556.
  12. Benhamed M, Herbig U, Ye T, Dejean A, Bischof O. Senescence is an endogenous trigger for microRNA-directed transcriptional gene silencing in human cells. Nat Cell Biol. 2012;14(3):266–275.
  13. Liang T, Guo L, Liu C. Genome-wide analysis of mir-548 gene family reveals evolutionary and functional implications. J Biomed Biotechnol. 2012;2012:679563.
  14. Zou L, Xu HG, Ren W, Jin R, Wang Y, Zhou GP. Transcriptional activation of the human CD2AP promoter by E2F1. PLoS One. 2012;7(8):e42774.
  15. Ni X, Liao Y, Li L, Zhang X, Wu Z. Therapeutic role of long non-coding RNA TCONS_00019174 in depressive disorders is dependent on Wnt/β-catenin signaling pathway. J Integr Neurosci. 2018;17(2):125–132. doi:10.3233/JIN-170052
  16. Gebeshuber CA, Kornauth C, Dong L, et al. Focal segmental glomerulosclerosis is induced by microRNA-193a and its downregulation of WT1. Nat Med. 2013;19(4):481–487.
  17. Luo Y, Wang C, Chen X, et al. Increased serum and urinary micro­RNAs in children with idiopathic nephrotic syndrome. Clin Chem. 2013;59(4):658–666.
  18. Wang G, Kwan BC, Lai FM, et al. Intrarenal expression of microRNAs in patients with IgA nephropathy. Lab Invest. 2010;90(1):98–103.
  19. Place RF, Li LC, Pookot D, Noonan EJ, Dahiya R. MicroRNA-373 induces expression of genes with complementary promoter sequences. Proc Natl Acad Sci U S A. 2008;105(5):1608–1613.
  20. Kim DH, Saetrom P, Snøve O Jr, Rossi JJ. MicroRNA-directed transcriptional gene silencing in mammalian cells. Proc Natl Acad Sci U S A. 2008;105(42):16230–16235.
  21. Younger ST, Corey DR. Transcriptional gene silencing in mammalian cells by miRNA mimics that target gene promoters. Nucleic Acids Res. 2011;39(13):5682–5691.
  22. Huang YP, Qiu LZ, Zhou GP. MicroRNA-939 downregulates CD2-associated protein by targeting promoter in HEK-293T cells. Ren Fail. 2016;38(4):508–513.