Transition Mutations in the hTERT Promoter Are Unrelated to Potential i-motif Formation in the C-Rich Strand

James Conrad, Mark L. Sowers, Dianne Y. Yap, Ellie Cherryhomes, B. Montgomery Pettitt, Kamil Khanipov, Lawrence C. Sowers

Research output: Contribution to journalArticlepeer-review

Abstract

Increased expression of the human telomere reverse transcriptase (hTERT) in tumors promotes tumor cell survival and diminishes the survival of patients. Cytosine-to-thymine (C-to-T) transition mutations (C250T or C228T) in the hTERT promoter create binding sites for transcription factors, which enhance transcription. The G-rich strand of the hTERT promoter can form G-quadruplex structures, whereas the C-rich strand can form an i-motif in which multiple cytosine residues are protonated. We considered the possibility that i-motif formation might promote cytosine deamination to uracil and C-to-T mutations. We computationally probed the accessibility of cytosine residues in an i-motif to attack by water. We experimentally examined regions of the C-rich strand to form i-motifs using pH-dependent UV and CD spectra. We then incubated the C-rich strand with and without the G-rich complementary strand DNA under various conditions, followed by deep sequencing. Surprisingly, deamination rates did not vary substantially across the 46 cytosines examined, and the two mutation hotspots were not deamination hotspots. The appearance of mutational hotspots in tumors is more likely the result of the selection of sequences with increased promoter binding affinity and hTERT expression.

Original languageEnglish (US)
Article number1308
JournalBiomolecules
Volume13
Issue number9
DOIs
StatePublished - Sep 2023

Keywords

  • cytosine deamination
  • quadruplex
  • telomerase
  • telomere
  • transition mutation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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