Cargando…
A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions
Cellular homeostasis requires the orderly expression of thousands of transcripts. Gene expression is regulated by numerous proteins that recognize post-translational modifications—in particular, the acetylation of lysine residues (Kac) on histones. In addition to affecting the general condensation s...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8984119/ https://www.ncbi.nlm.nih.gov/pubmed/35402505 http://dx.doi.org/10.3389/fmolb.2022.831758 |
_version_ | 1784682112384761856 |
---|---|
author | Loehr, Jérémy Kougnassoukou Tchara, Pata-Eting Gonthier, Kevin Noufi, Chahinez Linteau, Naomie Audet-Walsh, Étienne Lambert, Jean-Philippe |
author_facet | Loehr, Jérémy Kougnassoukou Tchara, Pata-Eting Gonthier, Kevin Noufi, Chahinez Linteau, Naomie Audet-Walsh, Étienne Lambert, Jean-Philippe |
author_sort | Loehr, Jérémy |
collection | PubMed |
description | Cellular homeostasis requires the orderly expression of thousands of transcripts. Gene expression is regulated by numerous proteins that recognize post-translational modifications—in particular, the acetylation of lysine residues (Kac) on histones. In addition to affecting the general condensation state of the chromatin, acetylated histones act as anchor points for bromodomain (BRD)-containing adapter proteins. BRDs are the primary Kac reader domains in humans, and proteins containing them act as chromatin scaffolds that organize large networks of interactions to regulate transcription. To characterize BRD-dependent interaction networks, we established cell lines in which histone acetylation is dependent on acetate supplementation. To do this, we used genome editing to knock out ATP citrate lyase (ACLY), the enzyme responsible for converting citrate to oxaloacetate and acetyl-CoA in the cytoplasm and nucleus. In our cellular model, removing acetate from the culture medium resulted in the rapid catabolism of acetylated histones to restore the nucleocytoplasmic acetyl-CoA pool. Here we report the use of our new model in functional proteomics studies to characterize BRD-dependent interaction networks on the chromatin. |
format | Online Article Text |
id | pubmed-8984119 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-89841192022-04-07 A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions Loehr, Jérémy Kougnassoukou Tchara, Pata-Eting Gonthier, Kevin Noufi, Chahinez Linteau, Naomie Audet-Walsh, Étienne Lambert, Jean-Philippe Front Mol Biosci Molecular Biosciences Cellular homeostasis requires the orderly expression of thousands of transcripts. Gene expression is regulated by numerous proteins that recognize post-translational modifications—in particular, the acetylation of lysine residues (Kac) on histones. In addition to affecting the general condensation state of the chromatin, acetylated histones act as anchor points for bromodomain (BRD)-containing adapter proteins. BRDs are the primary Kac reader domains in humans, and proteins containing them act as chromatin scaffolds that organize large networks of interactions to regulate transcription. To characterize BRD-dependent interaction networks, we established cell lines in which histone acetylation is dependent on acetate supplementation. To do this, we used genome editing to knock out ATP citrate lyase (ACLY), the enzyme responsible for converting citrate to oxaloacetate and acetyl-CoA in the cytoplasm and nucleus. In our cellular model, removing acetate from the culture medium resulted in the rapid catabolism of acetylated histones to restore the nucleocytoplasmic acetyl-CoA pool. Here we report the use of our new model in functional proteomics studies to characterize BRD-dependent interaction networks on the chromatin. Frontiers Media S.A. 2022-03-23 /pmc/articles/PMC8984119/ /pubmed/35402505 http://dx.doi.org/10.3389/fmolb.2022.831758 Text en Copyright © 2022 Loehr, Kougnassoukou Tchara, Gonthier, Noufi, Linteau, Audet-Walsh and Lambert. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Molecular Biosciences Loehr, Jérémy Kougnassoukou Tchara, Pata-Eting Gonthier, Kevin Noufi, Chahinez Linteau, Naomie Audet-Walsh, Étienne Lambert, Jean-Philippe A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title | A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title_full | A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title_fullStr | A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title_full_unstemmed | A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title_short | A Nutrient-Based Cellular Model to Characterize Acetylation-Dependent Protein-Protein Interactions |
title_sort | nutrient-based cellular model to characterize acetylation-dependent protein-protein interactions |
topic | Molecular Biosciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8984119/ https://www.ncbi.nlm.nih.gov/pubmed/35402505 http://dx.doi.org/10.3389/fmolb.2022.831758 |
work_keys_str_mv | AT loehrjeremy anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT kougnassoukoutcharapataeting anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT gonthierkevin anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT noufichahinez anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT linteaunaomie anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT audetwalshetienne anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT lambertjeanphilippe anutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT loehrjeremy nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT kougnassoukoutcharapataeting nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT gonthierkevin nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT noufichahinez nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT linteaunaomie nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT audetwalshetienne nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions AT lambertjeanphilippe nutrientbasedcellularmodeltocharacterizeacetylationdependentproteinproteininteractions |