Expression of metallothoinein isoform 3 is restricted at the post-transcriptional level in human bladder epithelial cells

Scott H. Garrett, Seongmi Park, Mary Ann Sens, Seema Somji, Rajendra K. Singh, Venugopal B R K Namburi, Donald A. Sens

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

This study was designed to define the effect that overexpression of MT-3 would have on a cell culture model of bladder urothelium. Stable and inducible transfection was used to achieve overexpression of the MT-3 gene in the UROtsa cell line. When the UROtsa cells were stably transfected with the MT-3 coding sequence, there was highly elevated expression of MT-3 mRNA, but no MT-3 protein. An inducible vector showed that low basal levels of MT-3 mRNA and protein could be produced, but that induction only increased MT-3 mRNA and not protein. The clones expressing low basal levels of MT-3 protein also had reduced growth rates compared to control cells. Site directed mutagenesis was used to produce an MT-3 coding sequence where the prolines in positions 7 and 9 were converted to threonines. When this altered MT-3 was stably transfected into the UROtsa cells, the cells were able to accumulate the mutated form of the MT-3 protein. These studies show that MT-3 protein expression is inhibited by post-transcriptional control in the urothelial cell. Modifying the MT-3 protein to resemble the MT-1 isoform removes this component of post-transcriptional control and allows accumulation of the mutated MT-3 protein. The altered sequence involved in post-transcriptional control of MT-3 protein expression is the same sequence implicated in the neuronal growth inhibitory activity associated specifically with the MT-3 isoform of the MT gene family.

Original languageEnglish (US)
Pages (from-to)66-74
Number of pages9
JournalToxicological Sciences
Volume87
Issue number1
DOIs
StatePublished - Sep 1 2005

Keywords

  • Bladder cancer
  • Cadmium
  • Growth inhibitory factor
  • Metallothionein
  • Post-transcriptional control
  • UROtsa
  • Urothelium

ASJC Scopus subject areas

  • Toxicology

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  • Cite this

    Garrett, S. H., Park, S., Sens, M. A., Somji, S., Singh, R. K., Namburi, V. B. R. K., & Sens, D. A. (2005). Expression of metallothoinein isoform 3 is restricted at the post-transcriptional level in human bladder epithelial cells. Toxicological Sciences, 87(1), 66-74. https://doi.org/10.1093/toxsci/kfi231