title: Identification of transcriptional and phosphatase regulators as interaction partners of human ADA3, a component of histone acetyltransferase complexes.
abstract: ADA (alteration/deficiency in activation) 3 is a conserved component of several transcriptional adaptor and HAT (histone acetyltransferase) complexes that regulate RNA polymerase II-mediated 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.
authors.affiliation: Department of Medical Biology, Faculty of Medicine, Pamukkale University, Denizli 20070, Turkey.
authors.forename: Sevil
authors.initials: S
authors.lastname: Zencir
authors.forename: Adam
authors.initials: A
authors.lastname: Sike
authors.forename: Melanie J
authors.initials: MJ
authors.lastname: Dobson
authors.forename: Ferhan
authors.initials: F
authors.lastname: Ayaydin
authors.forename: Imre
authors.initials: I
authors.lastname: Boros
authors.forename: Zeki
authors.initials: Z
authors.lastname: Topcu
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins; D017930:Genes, Reporter; D006367:HeLa Cells; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism*; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: C402637:AATF protein, human; D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C509751:PPP1R7 protein, human; C525691:PPP2R5D protein, human; D012097:Repressor Proteins; C097695:TADA2A protein, human; C444103:TADA3 protein, human; D014157:Transcription Factors; D051548:Histone Acetyltransferases; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2