Transcription factor Pit-1 affects transcriptional timing in the dual-promoter human prolactin gene

Anne V McNamara, Raheela Awais, Hiroshi Momiji, Lee Dunham, Karen Featherstone, Claire V Harper, Antony A Adamson, Sabrina Semprini, Nicholas A Jones, David G Spiller, John J Mullins, Bärbel F Finkenstädt, David Rand, Michael R H White, Julian R E Davis

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Abstract

Gene transcription occurs in short bursts interspersed with silent periods, and these kinetics can be altered by promoter structure. The effect of alternate promoter architecture on transcription bursting is not known. We studied the human prolactin (hPRL) gene that contains two promoters, a pituitary-specific promoter that requires the transcription factor Pit-1, and displays dramatic transcriptional bursting activity, and an alternate upstream promoter that is active in non-pituitary tissues. We studied large hPRL genomic fragments with luciferase reporters, and used bacterial artificial chromosome (BAC) recombineering to manipulate critical promoter regions. Stochastic switch mathematical modelling of single-cell time-lapse luminescence image data revealed that the Pit-1-dependent promoter showed longer, higher-amplitude transcriptional bursts. Knockdown studies confirmed that the presence of Pit-1 stabilised and prolonged periods of active transcription. Pit-1 therefore plays an active role in establishing the timing of transcription cycles, in addition to its cell-specific functions.

Original languageEnglish
JournalEndocrinology
Early online date3 Jan 2021
DOIs
Publication statusPublished - 3 Jan 2021

Keywords

  • Prolactin
  • transcription
  • pituitary
  • development
  • microscopy
  • prolactin gene
  • Pit-1
  • promoter
  • transcription timing
  • Gene transcription

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