Identification of transcriptional and phosphatase regulators as interaction partners of human ADA3, a component of histone acetyltransferase complexes

Sevil Zencir, Adam Sike, Melanie J. Dobson, F. Ayaydin, I. Boros, Zeki Topcu

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

ADA (alteration/deficiency in activation) 3 is a conserved component of several transcriptional adaptor and HAT (histone acetyltransferase) complexes that regulate RNA polymerase IImediated gene expression. Within the HAT complexes ADA3 is associated with ADA2 and the HAT GCN5 (general control non-repressed 5). ADA3 plays roles in diverse cellular processes and also in malignancies by modulating GCN5 catalytic activity and/or by interactions with other regulators. To gain a better understanding of ADA3 function, we used a yeast two-hybrid approach to screen a human fetal cDNA library for proteins that interacted with hADA3 (human ADA3). We identified three novel hADA3-interacting partners, a transcriptional regulator, AATF (apoptosis-antagonizing transcription factor), and regulatory subunits of the PP1 (protein phosphatase 1) and PP2A (protein phosphatase 2A) [PPP1R7 (PP1 regulatory subunit 7) and PPP2R5D (PP2A 56 kDa regulatory subunit δ isoform) respectively]. Analysis of truncated versions of hADA3 indicated that the C-terminal ADA2-interacting domain was not required for these interactions. Fluorescent microscopy analysis and co-immunoprecipitation provided support for the co-localization and interaction of hADA3 with these proteins in human cells. Expression of the interacting proteins altered expression of an hADA3-regulated reporter gene, suggesting functional consequences for the interactions. The detected interactions of hADA3 might extend the spectrum of mechanisms by which ADA3 can contribute to the regulation of gene expression and shed light on processes mediated by these newly identified ADA3 partners.

Original languageEnglish
Pages (from-to)311-320
Number of pages10
JournalBiochemical Journal
Volume450
Issue number2
DOIs
Publication statusPublished - Mar 1 2013

Fingerprint

Histone Acetyltransferases
Phosphoric Monoester Hydrolases
Protein Phosphatase 1
Gene expression
Protein Phosphatase 2
Proteins
DNA-Directed RNA Polymerases
Gene Library
Yeast
Catalyst activity
Microscopic examination
Protein Isoforms
Transcription Factors
Genes
Chemical activation
Cells
Apoptosis
Gene Expression Regulation
Reporter Genes
Immunoprecipitation

Keywords

  • Alteration/deficiency in activation 3 (ADA3)
  • Histone acetyltransferase
  • Protein-protein interaction
  • Spt/Ada/Gcn5/ acetyltransferase (SAGA)
  • Transcriptional regulation
  • Yeast two-hybrid technology

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Identification of transcriptional and phosphatase regulators as interaction partners of human ADA3, a component of histone acetyltransferase complexes. / Zencir, Sevil; Sike, Adam; Dobson, Melanie J.; Ayaydin, F.; Boros, I.; Topcu, Zeki.

In: Biochemical Journal, Vol. 450, No. 2, 01.03.2013, p. 311-320.

Research output: Contribution to journalArticle

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