Molecular Dissection of the Human Ubiquitin C Promoter Reveals Heat Shock Element Architectures with Activating and Repressive Functions

PloS One
Rita CrinelliMauro Magnani

Abstract

The promoter of the polyubiquitin C gene (UBC) contains putative heat shock elements (HSEs) which are thought to mediate UBC induction upon stress. However, the mapping and the functional characterization of the cis-acting determinants for its up-regulation have not yet been addressed. In this study, the sequence encompassing 916 nucleotides upstream of the transcription start site of the human UBC gene has been dissected by in silico, in vitro and in vivo approaches. The information derived from this analysis was used to study the functional role and the interplay of the identified HSEs in mediating the transcriptional activation of the UBC gene under conditions of proteotoxic stress, induced by the proteasome inhibitor MG132. Here we demonstrate that at least three HSEs, with different configurations, exist in the UBC promoter: two distal, residing within nucleotides -841/-817 and -715/-691, and one proximal to the transcription start site (nt -100/-65). All of them are bound by transcription factors belonging to the heat shock factor (HSF) family, as determined by bandshift, supershift and ChIP analyses. Site-directed mutagenesis of reporter constructs demonstrated that while the distal elements are involved in the up-regula...Continue Reading

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Citations

May 10, 2017·Omics : a Journal of Integrative Biology·Tolga AcunThomas Ried
Dec 29, 2016·Oncotarget·Verónica CánovasRosanna Paciucci
May 7, 2020·Applied Microbiology and Biotechnology·Tian-Yun Wang, Xiao Guo
Dec 19, 2018·Chemical Research in Toxicology·Annalida BediniGilberto Spadoni

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Methods Mentioned

BETA
ChIP-seq
ChIP
PCR
immunoprecipitation
transfection

Software Mentioned

TESS
MatInspector
TF search
SPSS
Primer Express

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