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

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McNamara, Anne V, Awais, Raheela, Momiji, Hiroshi, Dunham, Lee, Featherstone, Karen, Harper, Claire V, Adamson, Antony A, Semprini, Sabrina, Jones, Nicholas A, Spiller, David G, Mullins, John J, Finkenstädt, Bärbel, Rand, David A., White, Michael R. H. and Davis, Julian R. E. (2021) Transcription factor Pit-1 affects transcriptional timing in the dual-promoter human prolactin gene. Endocrinology, 162 (4). bqaa249. doi:10.1210/endocr/bqaa249 ISSN 0013-7227.

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Official URL: http://dx.doi.org/10.1210/endocr/bqaa249

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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.

Item Type: Journal Article
Subjects: Q Science > QA Mathematics
Q Science > QH Natural history
Q Science > QH Natural history > QH426 Genetics
Q Science > QP Physiology
Divisions: Faculty of Science, Engineering and Medicine > Science > Mathematics
Library of Congress Subject Headings (LCSH): Genetic transformation, Promoters (Genetics), Prolactin genes, Luciferases, Bacterial artificial chromosomes, Stochastic models
Journal or Publication Title: Endocrinology
Publisher: Endocrine Society
ISSN: 0013-7227
Official Date: April 2021
Dates:
DateEvent
April 2021Published
3 January 2021Available
29 December 2020Accepted
Volume: 162
Number: 4
Article Number: bqaa249
DOI: 10.1210/endocr/bqaa249
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 7 January 2021
Date of first compliant Open Access: 11 January 2021
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
091688Wellcome Trusthttp://dx.doi.org/10.13039/100010269
MR/K015885/1Medical Research Councilhttp://dx.doi.org/10.13039/501100000265
BB/K003097/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
EP/C544587/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
Studentship BB/F01760X/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
UNSPECIFIEDManchester Academic Health Sciences Centrehttps://healthinnovationmanchester.com/partnerships/manchester-academic-health-science-centre/

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