Revealing the alternative promoter usage of SAF/MAZ gene by bichromatic fluorescent reporter construct

Biosci Rep. 2019 Jan 18;39(1):BSR20171668. doi: 10.1042/BSR20171668. Print 2019 Jan 31.

Abstract

The large-scale identification of putative alternative promoters study shows more than 52% of human genes are regulated by alternative promoters. The human myc-associated zinc finger protein (SAF/MAZ) gene have SAF-1 and SAF-3 variants transcripted from two transcription start sites (TSSs). By using SAF/MAZ promoter as a model, we set up an approach to probe how the alternative promoters are regulated in real time. We have constructed the bichromatic fluorescent reporter driven by SAF/MAZ 5'-proximal promoter plasmids from which transactivation status of SAF-1 and SAF-3 alternative promoter could be monitored by EGFP and DsRed expression respectively. The results showed that the SAF-3 expression is regulated by alternative promoters. When the bichromatic fluorescent reporter was driven by -1692/+277 or -1401/+277 SAF/MAZ promoter the dominant expression of SAF-3 would be observed in comparison with SAF-1 expression. We also identified that Elk-1 is an inhibitory transcription factor for SAF-3 expression. The temporal diversity of SAF-1 and SAF-3 expressions can be observed via bichromatic fluorescent reporters. These imply that the bichromatic fluorescent reporter driven by alternative promoter construct might be a useful tool for decoding the temporal regulatory repertoire of alternative promoter in human genes.

Keywords: SAF-1; SAF-3; alternative promoter; bichromatic fluorescent reporter; myc-associated zinc finger protein gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions / genetics
  • Amino Acid Sequence / genetics
  • DNA-Binding Proteins / genetics*
  • Gene Expression Regulation / genetics*
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins / genetics
  • Humans
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Protein Isoforms / genetics*
  • Transcription Factors / genetics*
  • Transcription Initiation Site*
  • Transcriptional Activation / genetics
  • Transfection
  • ets-Domain Protein Elk-1 / genetics

Substances

  • 5' Untranslated Regions
  • DNA-Binding Proteins
  • Protein Isoforms
  • Transcription Factors
  • c-MYC-associated zinc finger protein
  • enhanced green fluorescent protein
  • ets-Domain Protein Elk-1
  • Green Fluorescent Proteins