A Natural Antisense Transcript Regulates Zeb2/Sip1 Gene Expression During Snail1-induced Epithelial-mesenchymal Transition

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2008 Mar 19

PMID 18347095

Citations 355

Authors

Manuel Beltran

Isabel Puig

Cristina Pena

Jose Miguel Garcia

Ana Belen Alvarez

Raul Pena

Felix Bonilla

Antonio Garcia de Herreros

Affiliations

Soon will be listed here.

Abstract

Expression of Snail1 in epithelial cells triggers an epithelial-mesenchymal transition (EMT). Here, we demonstrate that the synthesis of Zeb2, a transcriptional repressor of E-cadherin, is up-regulated after Snail1-induced EMT. Snail1 does not affect the synthesis of Zeb2 mRNA, but prevents the processing of a large intron located in its 5'-untranslated region (UTR). This intron contains an internal ribosome entry site (IRES) necessary for the expression of Zeb2. Maintenance of 5'-UTR Zeb2 intron is dependent on the expression of a natural antisense transcript (NAT) that overlaps the 5' splice site in the intron. Ectopic overexpression of this NAT in epithelial cells prevents splicing of the Zeb2 5'-UTR, increases the levels of Zeb2 protein, and consequently down-regulates E-cadherin mRNA and protein. The relevance of these results is demonstrated by the strong association between NAT presence and conservation of the 5'-UTR intron in cells that have undergone EMT or in human tumors with low E-cadherin expression. Therefore, the results presented in this article reveal the existence of a NAT capable of activating Zeb2 expression, explain the mechanism involved in this activation, and demonstrate that this NAT regulates E-cadherin expression.

Citing Articles

ZEB2 Gene Pathogenic Variants Across Protein-Coding Regions and Impact on Clinical Manifestations: A Review.

Hossain W, St Peter C, Lovell S, Rafi S, Butler M Int J Mol Sci. 2025; 26(3).

PMID: 39941075 PMC: 11818587. DOI: 10.3390/ijms26031307.

CDH3-AS1 antisense RNA enhances P-cadherin translation and acts as a tumor suppressor in melanoma.

Chadourne M, Griffith C, Xu X, Brennan E, Vera O, Mecozzi N bioRxiv. 2025; .

PMID: 39764055 PMC: 11703152. DOI: 10.1101/2024.12.26.630428.

Computational Resources for lncRNA Functions and Targetome.

Thakur A, Kumar M Methods Mol Biol. 2024; 2883:299-323.

PMID: 39702714 DOI: 10.1007/978-1-0716-4290-0_13.

Novel insights on the positive correlation between sense and antisense pairs on gene expression.

Das S, Zea Rojas M, Tran E Wiley Interdiscip Rev RNA. 2024; 15(4):e1864.

PMID: 39087253 PMC: 11626863. DOI: 10.1002/wrna.1864.

Hypoxia-inducible factor 3α1 increases epithelial-to-mesenchymal transition and iron uptake to drive colorectal cancer liver metastasis.

Villareal L, Falcon D, Xie L, Xue X Br J Cancer. 2024; 130(12):1904-1915.

PMID: 38693428 PMC: 11183190. DOI: 10.1038/s41416-024-02699-3.

References

Dahary D, Elroy-Stein O, Sorek R . Naturally occurring antisense: transcriptional leakage or real overlap?. Genome Res. 2005; 15(3):364-8. PMC: 551562. DOI: 10.1101/gr.3308405. View

Comijn J, Berx G, Vermassen P, Verschueren K, van Grunsven L, Bruyneel E . The two-handed E box binding zinc finger protein SIP1 downregulates E-cadherin and induces invasion. Mol Cell. 2001; 7(6):1267-78. DOI: 10.1016/s1097-2765(01)00260-x. View

Long J, Zuo D, Park M . Pc2-mediated sumoylation of Smad-interacting protein 1 attenuates transcriptional repression of E-cadherin. J Biol Chem. 2005; 280(42):35477-89. DOI: 10.1074/jbc.M504477200. View

Barbera M, Puig I, Dominguez D, Julien-Grille S, Guaita-Esteruelas S, Peiro S . Regulation of Snail transcription during epithelial to mesenchymal transition of tumor cells. Oncogene. 2004; 23(44):7345-54. DOI: 10.1038/sj.onc.1207990. View

Prescott E, Proudfoot N . Transcriptional collision between convergent genes in budding yeast. Proc Natl Acad Sci U S A. 2002; 99(13):8796-801. PMC: 124378. DOI: 10.1073/pnas.132270899. View

Hellen C, Sarnow P . Internal ribosome entry sites in eukaryotic mRNA molecules. Genes Dev. 2001; 15(13):1593-612. DOI: 10.1101/gad.891101. View

Verschueren K, Remacle J, Collart C, Kraft H, Baker B, Tylzanowski P . SIP1, a novel zinc finger/homeodomain repressor, interacts with Smad proteins and binds to 5'-CACCT sequences in candidate target genes. J Biol Chem. 1999; 274(29):20489-98. DOI: 10.1074/jbc.274.29.20489. View

Esteban M, Tran M, Harten S, Hill P, Castellanos M, Chandra A . Regulation of E-cadherin expression by VHL and hypoxia-inducible factor. Cancer Res. 2006; 66(7):3567-75. DOI: 10.1158/0008-5472.CAN-05-2670. View

Hastings M, Ingle H, Lazar M, Munroe S . Post-transcriptional regulation of thyroid hormone receptor expression by cis-acting sequences and a naturally occurring antisense RNA. J Biol Chem. 2001; 275(15):11507-13. DOI: 10.1074/jbc.275.15.11507. View

10.

Rougeulle C, Heard E . Antisense RNA in imprinting: spreading silence through Air. Trends Genet. 2002; 18(9):434-7. DOI: 10.1016/s0168-9525(02)02749-x. View

11.

Eger A, Aigner K, Sonderegger S, Dampier B, Oehler S, Schreiber M . DeltaEF1 is a transcriptional repressor of E-cadherin and regulates epithelial plasticity in breast cancer cells. Oncogene. 2005; 24(14):2375-85. DOI: 10.1038/sj.onc.1208429. View

12.

Batlle E, Verdu J, Dominguez D, del Mont Llosas M, Diaz V, Loukili N . Protein kinase C-alpha activity inversely modulates invasion and growth of intestinal cells. J Biol Chem. 1998; 273(24):15091-8. DOI: 10.1074/jbc.273.24.15091. View

13.

Peters N, Rohrbach J, Zalewski B, Byrkett C, Vaughn J . RNA editing and regulation of Drosophila 4f-rnp expression by sas-10 antisense readthrough mRNA transcripts. RNA. 2003; 9(6):698-710. PMC: 1370437. DOI: 10.1261/rna.2120703. View

14.

Lehner B, Williams G, Duncan Campbell R, Sanderson C . Antisense transcripts in the human genome. Trends Genet. 2002; 18(2):63-5. DOI: 10.1016/s0168-9525(02)02598-2. View

15.

Evans A, Russell R, Roche O, Burry T, Fish J, Chow V . VHL promotes E2 box-dependent E-cadherin transcription by HIF-mediated regulation of SIP1 and snail. Mol Cell Biol. 2006; 27(1):157-69. PMC: 1800649. DOI: 10.1128/MCB.00892-06. View

16.

Peinado H, Olmeda D, Cano A . Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype?. Nat Rev Cancer. 2007; 7(6):415-28. DOI: 10.1038/nrc2131. View

17.

Krishnamachary B, Zagzag D, Nagasawa H, Rainey K, Okuyama H, Baek J . Hypoxia-inducible factor-1-dependent repression of E-cadherin in von Hippel-Lindau tumor suppressor-null renal cell carcinoma mediated by TCF3, ZFHX1A, and ZFHX1B. Cancer Res. 2006; 66(5):2725-31. DOI: 10.1158/0008-5472.CAN-05-3719. View

18.

Batlle E, Sancho E, Franci C, Dominguez D, Monfar M, Baulida J . The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol. 2000; 2(2):84-9. DOI: 10.1038/35000034. View

19.

Katayama S, Tomaru Y, Kasukawa T, Waki K, Nakanishi M, Nakamura M . Antisense transcription in the mammalian transcriptome. Science. 2005; 309(5740):1564-6. DOI: 10.1126/science.1112009. View

20.

Vandewalle C, Comijn J, De Craene B, Vermassen P, Bruyneel E, Andersen H . SIP1/ZEB2 induces EMT by repressing genes of different epithelial cell-cell junctions. Nucleic Acids Res. 2005; 33(20):6566-78. PMC: 1298926. DOI: 10.1093/nar/gki965. View