Modulation of STAT1-Driven Transcriptional Activity by Histone Deacetylases

Methods Mol Biol. 2017;1510:277-285. doi: 10.1007/978-1-4939-6527-4_20.

Abstract

The luciferase (LUC) reporter assay is commonly used to study gene expression at the transcriptional level. It is convenient, fast, sensitive, inexpensive, and provides quantitative data about small changes in transcription. Signal transducer and activator of transcription 1 (STAT1) is a transcription factor that plays a crucial role in signaling by interferons (IFNs). Here, we describe LUC reporter studies that address the role of histone deacetylase (HDAC) activity in STAT1-dependent gene activation. These experiments include overexpression of HDAC1, HDAC2, HDAC3, and HDAC4 as well as silencing of HDAC1, HDAC2, and HDAC3 through RNA interference in mammalian cancer cells.

Keywords: HDAC; HDAC inhibitors; Interferons; Luciferase gene reporter; STAT1.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Biological Assay*
  • Butyric Acid / pharmacology
  • Cell Line, Tumor
  • Epigenesis, Genetic
  • Genes, Reporter
  • HCT116 Cells
  • Histone Deacetylase 1 / antagonists & inhibitors
  • Histone Deacetylase 1 / genetics*
  • Histone Deacetylase 1 / metabolism
  • Histone Deacetylase 2 / antagonists & inhibitors
  • Histone Deacetylase 2 / genetics*
  • Histone Deacetylase 2 / metabolism
  • Histone Deacetylase Inhibitors / pharmacology*
  • Histone Deacetylases / genetics
  • Histone Deacetylases / metabolism
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Interferon-gamma / pharmacology
  • Luciferases / genetics
  • Luciferases / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • STAT1 Transcription Factor / genetics*
  • STAT1 Transcription Factor / metabolism
  • Transcription, Genetic / drug effects*

Substances

  • Histone Deacetylase Inhibitors
  • Histones
  • RNA, Small Interfering
  • Repressor Proteins
  • STAT1 Transcription Factor
  • STAT1 protein, human
  • Butyric Acid
  • Interferon-gamma
  • Luciferases
  • HDAC1 protein, human
  • HDAC2 protein, human
  • HDAC4 protein, human
  • Histone Deacetylase 1
  • Histone Deacetylase 2
  • Histone Deacetylases
  • histone deacetylase 3