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Differential Translational Regulation of SLUG mRNA By a uORF In SLUG‐High and SLUG‐Low Cancer Cells
Author(s) -
Yarlagadda Vidhush K,
Misra Smita,
Mittal Mukul K,
Chaudhuri Gautam
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb62
Subject(s) - slug , upstream open reading frame , five prime untranslated region , untranslated region , biology , luciferase , messenger rna , three prime untranslated region , microbiology and biotechnology , open reading frame , gene , cancer research , transfection , genetics , peptide sequence
The metastasis regulator protein SLUG is stringently regulated during cellular growth and development. This gene is expressed from a single promoter and produces an mRNA with a 164 nucleotide 5′‐untranslated region (5′‐UTR). There is a highly conserved tricodon upstream open reading frame (uORF) at the 5′‐UTR in human SLUG mRNA. uORFs are known to regulate the translation of mRNAs. Here we report differential inhibition of SLUG mRNA translation by this uORF in SLUG‐high and SLUG‐low breast cancer cells. We cloned the 164 base‐pair 5′‐UTR sequence in front of the firefly luciferase gene in pGL3‐Control plasmid in the appropriate orientation. Dual luciferase assay with transiently transfected cells showed 50–60% inhibition of the expression of luciferase activity in the SLUG‐low but not in the SLUG‐high breast cancer cells. Mutagenesis of the ATG codon in the uORF to TAG abrogated this inhibition. This data indicates that human SLUG gene expression is differentially regulated by the uORF. We are currently investigating the mechanism behind this differential regulation of SLUG expression. Understanding the molecular mechanism of this inhibition will contribute towards the development of chemical intervention measures against the expression of this important metastasis regulatory protein and thus against breast cancer metastasis.