Gene-specific transcriptional activation mediated by the p150 subunit of the chromatin assembly factor 1

Sung-Bau Lee, Derick S C Ou, Chung-Fan Lee, Li-Jung Juan

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Chromatin assembly factor 1 contains three subunits, p150, p60, and p48. It is essential for coupling nucleosome assembly to newly synthesized DNA. Whether chromatin assembly factor 1 subunits have functions beyond escorting histones, which depends on the complex formation of p150 and p60, has been an issue of great interest. This study reveals a novel role of p150, but not p60, in gene-specific transcriptional activation. We found that p150 transcriptionally activated an essential viral promoter, the major immediate early promoter (MIEP) of the human cytomegalovirus, independently of p60. Knocking down p150 decreased the MIEP function in both transfected and virally infected cells. The chromatin immunoprecipitation analysis and the in vitro protein-DNA binding assay demonstrated that p150 used its KER domain to associate with the MIEP from -593 to -574 bp. The N-terminal 244 residues were also found essential for p150-mediated MIEP activation, likely through recruiting the acetyltransferase p300 to acetylate local histones. Domain swapping experiments further showed that the KER and theNterminus of p150 acted as an independent DNA binding and transcriptional activation domain, respectively. Because p60 did not seem involved in the reaction, together these results indicate for the first time that p150 directly activates transcription, independently of its histone deposition function. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
Original languageEnglish
Pages (from-to)14040-14049
Number of pages10
JournalJournal of Biological Chemistry
Volume284
Issue number21
DOIs
Publication statusPublished - 2009
Externally publishedYes

    Fingerprint

Keywords

  • Acetyl transferase
  • Chromatin assembly
  • Chromatin immunoprecipitation analysis
  • Complex formations
  • DNA binding
  • Domain swapping
  • Human cytomegalovirus
  • In-vitro
  • Infected cells
  • N-terminal
  • Nucleosome assembly
  • Protein-DNA binding
  • Transcriptional activation domain
  • Transcriptional activations
  • Viral promoters
  • Genes
  • Nucleic acids
  • Transcription
  • DNA
  • acyltransferase
  • acyltransferase p300
  • arginine
  • chromatin assembly factor 1
  • chromatin assembly factor 1 subunit p150
  • chromatin assembly factor 1 subunit p60
  • glutamic acid
  • histone
  • lysine
  • unclassified drug
  • chromatin assembly factor I
  • DNA binding protein
  • histone acetyltransferase PCAF
  • immediate early protein
  • nonhistone protein
  • p300-CBP-associated factor
  • protein subunit
  • amino terminal sequence
  • article
  • chromatin assembly and disassembly
  • chromatin immunoprecipitation
  • controlled study
  • cytomegalovirus infection
  • human
  • human cell
  • in vitro study
  • major immediate early promoter
  • priority journal
  • promoter region
  • protein DNA binding
  • protein domain
  • protein function
  • transcription initiation
  • base pairing
  • chemistry
  • Cytomegalovirus
  • gene expression regulation
  • genetic transfection
  • genetics
  • metabolism
  • protein binding
  • protein tertiary structure
  • tumor cell line
  • Human herpesvirus 5
  • Base Pairing
  • Cell Line, Tumor
  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Gene Expression Regulation, Viral
  • Humans
  • Immediate-Early Proteins
  • p300-CBP Transcription Factors
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Subunits
  • Transcriptional Activation
  • Transfection

Cite this