Short stretches of rare codons regulate translation of the transcription factor ZEB2 in cancer cells

Abstract

Two proteins comprising the ZEB family of zinc finger transcription factors, ZEB1 and ZEB2, execute EMT programs in embryonic development and cancer. By studying regulation of their expression, we describe a novel mechanism that limits ZEB2 protein synthesis. A protein motif located at the border of the SMAD-binding domain of ZEB2 protein induces ribosomal pausing and compromises protein synthesis. The function of this protein motif is dependent on stretches of rare codons, Leu(UUA)-Gly(GGU)-Val(GUA). Incorporation of these triplets in the homologous region of ZEB1 does not affect protein translation. Our data suggest that rare codons have a regulatory role only if they are present within appropriate protein structures. We speculate that pools of transfer RNA available for protein translation impact on the configuration of epithelial mesenchymal transition pathways in tumor cells.

Long non‐coding RNA ZEB2‐AS1 promotes the proliferation, metastasis and epithelial mesenchymal transition in triple‐negative breast cancer by epigenetically activating ZEB2

“…The proliferation capacities of the cells cultured for 24,48,72,96, and 120 hours were tested by the CCK-8 assays. The proliferation capacities of the cells cultured for 24,48,72,96, and 120 hours were tested by the CCK-8 assays.…”

“…Transcription factor attributing to especial Zinc finger protein families, usually located in the downstream of some signaling pathways executing EMT process in normal and pathological conditions 24. A, Tumor volume growth curves in SCID mice.…”

Modulation of HIV-1 Gag/Gag-Pol frameshifting by tRNA abundance

“…Cellular tRNA levels for tRNA UAA Leu , tRNA CAG Leu , tRNA UAC Val and tRNA CAC Val were quantified using a strategy published by (53). Total cellular RNA was extracted using the RNeasy Mini Kit (Qiagen) according to the manufacturer's protocol.…”

“…To eliminate potentially co-isolated DNA from the samples, 0.5 g RNA was incubated with DNase I (New England Biolabs) in a final volume of 10 l for 30 min at 37 • C and finally heated to 65 • C for 5 min to inactivate the enzyme. The primer for the reverse transcription entailed a tRNA-specific sequence and a stem-loop sequence (53). The tRNA-specific primer was the same for tRNA UAA Leu and tRNA CAG Leu (complementary to positions [47][48][49][50][51][52][53][54][55] (53); the forward primer for the qPCR is highly specific to each tRNA and the reverse primer is universal ( Supplementary Table S5).…”

“…Error bars represent S.E.M of three biological replicates with three technical replicates each. Experimental design of qRT-PCR was according to (53). Human cell lines are indicated below the graph.…”