Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.

We spliced the promoters of the human telomerase and human survivin genes (PhTERT and PhSurv, respectively) widely used for gene therapy and known to have the broadest cancer type spectrum of activity. Two head-to-tail constructs were obtained: the PhTERT-PhSurv and PhSurv-PhTERT tandems. The splici...

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Autores principales:	Irina V Alekseenko, Victor V Pleshkan, Eugene P Kopantzev, Elena A Stukacheva, Igor P Chernov, Tatyana V Vinogradova, Eugene D Sverdlov
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fc252ac0142c49f6a02ce405beb181d2
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spelling	oai:doaj.org-article:fc252ac0142c49f6a02ce405beb181d22021-11-18T08:13:21ZActivity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.1932-620310.1371/journal.pone.0046474https://doaj.org/article/fc252ac0142c49f6a02ce405beb181d22012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23056318/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203We spliced the promoters of the human telomerase and human survivin genes (PhTERT and PhSurv, respectively) widely used for gene therapy and known to have the broadest cancer type spectrum of activity. Two head-to-tail constructs were obtained: the PhTERT-PhSurv and PhSurv-PhTERT tandems. The splicing caused quantitative and qualitative changes in the promoter features. In both constructs, only the promoter proximal to the transcribed gene retained its ability to initiate transcription, whereas the distal promoter was silent, the phenomenon never reported before. However, the distal promoter modulated the activity of the proximal one by increasing its strength and causing an appearance of additional transcription start sites. We suggested that this suppression might be due to the presence of Sp1 transcription factor binding sites in both promoters and Sp1-bridges between these sites. Such Sp1-bridges might convert the tandem promoter linear DNA into a stem-loop structure. If localized inside the formed loop, the distal promoter could lose its ability to initiate transcription. To test this hypothesis, we constructed two modified double promoters, where the proximal PhSurv promoter was replaced either by a shortened variant of the survivin promoter (PhSurv269) or by the mouse survivin promoter. Both PhSurv substitutes were considerably shorter than PhSurv and had different numbers and/or positions of Sp1 sites. In modified tandems, transcription was initiated from both promoters. We also prepared two mutant forms of the PhSurv-PhTERT tandem with two or four Sp1 sites removed from the distal "long" PhSurv promoter. In the first case, the distal PhSurv promoter remained silent, whereas the removal of four Sp1 binding sites restored its activity. In the majority of studied cancer cell lines the efficiency of transcription from the hTERT-(shortened hSurv269) promoter tandem was markedly higher than from each constituent promoter. In normal lung fibroblast cells, the tandem promoter activity was considerably lower.Irina V AlekseenkoVictor V PleshkanEugene P KopantzevElena A StukachevaIgor P ChernovTatyana V VinogradovaEugene D SverdlovPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 10, p e46474 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Irina V Alekseenko Victor V Pleshkan Eugene P Kopantzev Elena A Stukacheva Igor P Chernov Tatyana V Vinogradova Eugene D Sverdlov Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
description	We spliced the promoters of the human telomerase and human survivin genes (PhTERT and PhSurv, respectively) widely used for gene therapy and known to have the broadest cancer type spectrum of activity. Two head-to-tail constructs were obtained: the PhTERT-PhSurv and PhSurv-PhTERT tandems. The splicing caused quantitative and qualitative changes in the promoter features. In both constructs, only the promoter proximal to the transcribed gene retained its ability to initiate transcription, whereas the distal promoter was silent, the phenomenon never reported before. However, the distal promoter modulated the activity of the proximal one by increasing its strength and causing an appearance of additional transcription start sites. We suggested that this suppression might be due to the presence of Sp1 transcription factor binding sites in both promoters and Sp1-bridges between these sites. Such Sp1-bridges might convert the tandem promoter linear DNA into a stem-loop structure. If localized inside the formed loop, the distal promoter could lose its ability to initiate transcription. To test this hypothesis, we constructed two modified double promoters, where the proximal PhSurv promoter was replaced either by a shortened variant of the survivin promoter (PhSurv269) or by the mouse survivin promoter. Both PhSurv substitutes were considerably shorter than PhSurv and had different numbers and/or positions of Sp1 sites. In modified tandems, transcription was initiated from both promoters. We also prepared two mutant forms of the PhSurv-PhTERT tandem with two or four Sp1 sites removed from the distal "long" PhSurv promoter. In the first case, the distal PhSurv promoter remained silent, whereas the removal of four Sp1 binding sites restored its activity. In the majority of studied cancer cell lines the efficiency of transcription from the hTERT-(shortened hSurv269) promoter tandem was markedly higher than from each constituent promoter. In normal lung fibroblast cells, the tandem promoter activity was considerably lower.
format	article
author	Irina V Alekseenko Victor V Pleshkan Eugene P Kopantzev Elena A Stukacheva Igor P Chernov Tatyana V Vinogradova Eugene D Sverdlov
author_facet	Irina V Alekseenko Victor V Pleshkan Eugene P Kopantzev Elena A Stukacheva Igor P Chernov Tatyana V Vinogradova Eugene D Sverdlov
author_sort	Irina V Alekseenko
title	Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
title_short	Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
title_full	Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
title_fullStr	Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
title_full_unstemmed	Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.
title_sort	activity of the upstream component of tandem tert/survivin promoters depends on features of the downstream component.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/fc252ac0142c49f6a02ce405beb181d2
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Activity of the upstream component of tandem TERT/survivin promoters depends on features of the downstream component.

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