PITX1 protein interacts with ZCCHC10 to regulate hTERT mRNA transcription

PLoS One. 2019 Aug 12;14(8):e0217605. doi: 10.1371/journal.pone.0217605. eCollection 2019.

Abstract

Telomerase is a ribonucleoprotein ribonucleic enzyme that is essential for cellular immortalization via elongation of telomere repeat sequences at the end of chromosomes. Human telomerase reverse transcriptase (hTERT), the catalytic subunit of telomerase holoenzyme, is a key regulator of telomerase activity. Telomerase activity, which has been detected in the majority of cancer cells, is accompanied by hTERT expression, suggesting that this enzyme activity contributes to an unlimited replication potential of cancer cells via regulation of telomere length. Thus, hTERT is an attractive target for cancer-specific treatments. We previously reported that pared-like homeodomain 1 (PITX1) is a negative regulator of hTERT through direct binding to the hTERT promoter. However, the mechanism by which the function of PITX1 contributes to transcriptional silencing of the hTERT gene remains to be clarified. Here, we show that PITX1 and zinc finger CCHC-type containing 10 (ZCCHC10) proteins cooperate to facilitate the transcriptional regulation of the hTERT gene by functional studies via FLAG pull-down assay. Co-expression of PITX1 and ZCCHC10 resulted in inhibition of hTERT transcription, in melanoma cell lines, whereas mutate-deletion of homeodomain in PITX1 that interact with ZCCHC10 did not induce similar phenotypes. In addition, ZCCHC10 expression levels showed marked decrease in the majority of melanoma cell lines and tissues. Taken together, these results suggest that ZCCHC10-PITX1 complex is the functional unit that suppresses hTERT transcription, and may play a crucial role as a novel tumor suppressor complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation, Enzymologic*
  • Humans
  • Melanoma / genetics
  • Melanoma / metabolism*
  • Paired Box Transcription Factors / genetics
  • Paired Box Transcription Factors / metabolism*
  • Promoter Regions, Genetic
  • Protein Interaction Domains and Motifs
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Telomerase / genetics
  • Telomerase / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • Tumor Cells, Cultured
  • Zinc Fingers / genetics*

Substances

  • Paired Box Transcription Factors
  • RNA, Messenger
  • Transcription Factors
  • homeobox protein PITX1
  • TERT protein, human
  • Telomerase

Grants and funding

This work was supported by Japan Society for the Promotion of Science (JSPS), a Grant-in-Aid for Young Scientists (Start-up) Grant Number 15H06414, Early-Career Scientists Grant Number 18K16030, and Grant-in-Aid for Scientific Research (C) Grant Number 15K08304. This research was partly performed at the Tottori Bio Frontier managed by Tottori prefecture. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.