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Related Experiment Videos

Switch from Myc/Max to Mad1/Max binding and decrease in histone acetylation at the telomerase reverse transcriptase

D Xu1, N Popov, M Hou

  • 1Department of Medicine, Division of Hematology, Karolinska Hospital, SE-171 76 Stockholm, Sweden. Dawei.Xu@cmm.ki.se

Proceedings of the National Academy of Sciences of the United States of America
|March 29, 2001
PubMed
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Myc and Mad1 proteins regulate human telomerase reverse transcriptase (hTERT) gene expression. Reciprocal binding to the hTERT promoter controls gene activation and repression during cell differentiation, involving histone acetylation.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Epigenetics

Background:

  • The human telomerase reverse transcriptase (hTERT) gene encodes the catalytic subunit of telomerase, crucial for cell proliferation.
  • Myc and Mad1 proteins are implicated in regulating hTERT expression, but their precise roles and interactions at the promoter are not fully understood.

Purpose of the Study:

  • To investigate the in vivo interaction of c-Myc and Mad1 proteins with the hTERT promoter in HL60 cells.
  • To elucidate the role of these proteins and histone acetylation in regulating hTERT transcription during cell differentiation.

Main Methods:

  • Chromatin immunoprecipitation (ChIP) assay to analyze in vivo protein-DNA interactions.
  • Treatment with DMSO for cell differentiation and trichostatin A (a histone deacetylase inhibitor).

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Main Results:

  • c-Myc occupied the hTERT promoter in proliferating HL60 cells, while Mad1 occupied it in differentiated cells.
  • Histone acetylation at the hTERT promoter decreased upon differentiation.
  • Trichostatin A inhibited histone deacetylation, attenuated hTERT repression during differentiation, and activated hTERT transcription in resting cells.

Conclusions:

  • Reciprocal E-box occupancy by c-Myc and Mad1 regulates hTERT gene activation and repression.
  • Histone acetylation/deacetylation plays a critical role in controlling hTERT expression during cell differentiation and in various cell types.