trans Repression of the Human Metallothionein IIA Gene Promoter by PZ120, a Novel 120-Kilodalton Zinc Finger Protein

Abstract

Metallothioneins are small, highly conserved, cysteine-rich proteins that bind a variety of metal ions. They are found in virtually all eukaryotic organisms and are regulated primarily at the transcriptional level. In humans, the predominant metallothionein gene is hMTIIA, which accounts for 50% of all metallothioneins expressed in cultured human cells. The hMTIIA promoter is quite complex. In addition to cis-acting DNA sequences that serve as binding sites for trans-acting factors such as Sp1, AP1, AP2, AP4, and the glucocorticoid receptor, the hMTIIA promoter contains eight consensus metal response element sequences. We report here the cloning of a novel zinc finger protein with a molecular mass of 120 kDa (PZ120) that interacts specifically with the hMTIIA transcription initiation site. The PZ120 protein is ubiquitously expressed in most tissues and possesses a conserved poxvirus and zinc finger (POZ) motif previously found in several zinc finger transcription factors. Intriguingly, we found that a region of PZ120 outside of the zinc finger domain can bind specifically to the hMTIIA DNA. Using transient-transfection analysis, we found that PZ120 repressed transcription of the hMTIIA promoter. These results suggest that the hMTIIA gene is regulated by an additional negative regulator that has not been previously described.

ACKNOWLEDGMENTS

We thank Alan Tomkinson for purified RAD1 protein; Mon-Li Chu for the cDNA library; Nikola Valkov for assistance with microscopy; Tom Shenk, Rosalind Jackson, Eden Kahle, and Julia Lee for discussion and critical reading of the manuscript; and the Moffitt Cancer Center Imaging Facility for technical support.

This work was supported by a grant from the National Institutes of Health (ES09262) to E.S.