Gawky modulates MTF-1-mediated transcription activation and metal discrimination

Abstract

Metal-induced genes are usually transcribed at relatively low levels under normal conditions and are rapidly activated by heavy metal stress. Many of these genes respond preferentially to specific metal-stressed conditions. However, the mechanism by which the general transcription machinery discriminates metal stress from normal conditions and the regulation of MTF-1-meditated metal discrimination are poorly characterized. Using a focused RNAi screening in Drosophila Schneider 2 (S2) cells, we identified a novel activator, the Drosophila gawky, of metal-responsive genes. Depletion of gawky has almost no effect on the basal transcription of the metallothionein (MT) genes, but impairs the metal-induced transcription by inducing the dissociation of MTF-1 from the MT promoters and the deficient nuclear import of MTF-1 under metal-stressed conditions. This suggests that gawky serves as a ‘checkpoint’ for metal stress and metal-induced transcription. In fact, regular mRNAs are converted into gawky-controlled transcripts if expressed under the control of a metal-responsive promoter, suggesting that whether transcription undergoes gawky-mediated regulation is encrypted therein. Additionally, lack of gawky eliminates the DNA binding bias of MTF-1 and the transcription preference of metal-specific genes. This suggests a combinatorial control of metal discrimination by gawky, MTF-1, and MTF-1 binding sites.

Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae

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Transcriptional Regulation and Protein Localization of Zip10, Zip13 and Zip14 Transporters of Freshwater Teleost Yellow Catfish Pelteobagrus fulvidraco Following Zn Exposure in a Heterologous HEK293T Model

“…Most TF binding sites are a 4 bp- to 30-bp wide integral DNA sequence [ 62 ]. Furthermore, the core consensus sequence of the MRE is TGCRCNC, in which R represents A or G and N represents any nucleotide [ 63 ]. STAT3-binding sites are characterized by clusters of conserved motifs with a core sequence of TTCT/CNA/GGAA, in which N represents any nucleotide [ 64 ].…”

Exportin 4 depletion leads to nuclear accumulation of a subset of circular RNAs

“…For overexpression with Exportins, eIF5A, Smad3, eIF4E, Rpa1, Ddx39a, Ddx39b C-terminal FLAG-tag transfection in this study, sequence of the target was cloned into the p3xflag-myc-cmv-24 between HindIII and BamHI sites. To generate the plasmid expressing FLAG-tagged Ranbp16 (Hy_pAct5C FLAG-Ranbp16), the Ranbp16 CDS was inserted into the MCS site of our previously-described Hy_pAct5C Flag MCS plasmid 78 . The shRNA plasmid for knockdown of eIF5A mRNA (sheIF5A, TRCN0000010985), RPA1 mRNA (shRPA1, TRCN0000071282), SRSF1 mRNA (shSRSF1, TRCN0000109143), SF3A1 mRNA (shSF3A1, TRCN0000109205), SF3B1 mRNA (shSF3B1, TRCN0000123809) with negative-control shRNA (shC002) was obtained from the MISSION shRNA Library (Sigma-Aldrich, Germany).…”