The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast

Intracellular trafficking pathways control residency and bioactivity of integral membrane proteins at the cell surface. Upon internalisation, surface cargo proteins can be delivered back to the plasma membrane via endosomal recycling pathways. Recycling is thought to be controlled at the metabolic a...

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Autores principales:	Konstantina Amoiradaki, Kate R. Bunting, Katherine M. Paine, Josephine E. Ayre, Karen Hogg, Kamilla M. E. Laidlaw, Chris MacDonald
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	endocytosis membrane trafficking surface membrane proteins histone deacetylase transcription Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/ea2cd7d020234a359aaee50b401a06dc
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spelling	oai:doaj.org-article:ea2cd7d020234a359aaee50b401a06dc2021-11-25T17:57:00ZThe Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast10.3390/ijms2222124771422-00671661-6596https://doaj.org/article/ea2cd7d020234a359aaee50b401a06dc2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12477https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Intracellular trafficking pathways control residency and bioactivity of integral membrane proteins at the cell surface. Upon internalisation, surface cargo proteins can be delivered back to the plasma membrane via endosomal recycling pathways. Recycling is thought to be controlled at the metabolic and transcriptional level, but such mechanisms are not fully understood. In yeast, recycling of surface proteins can be triggered by cargo deubiquitination and a series of molecular factors have been implicated in this trafficking. In this study, we follow up on the observation that many subunits of the Rpd3 lysine deacetylase complex are required for recycling. We validate ten Rpd3-complex subunits in recycling using two distinct assays and developed tools to quantify both. Fluorescently labelled Rpd3 localises to the nucleus and complements recycling defects, which we hypothesised were mediated by modulated expression of Rpd3 target gene(s). Bioinformatics implicated 32 candidates that function downstream of Rpd3, which were over-expressed and assessed for capacity to suppress recycling defects of <i>rpd3</i>∆ cells. This effort yielded three hits: Sit4, Dit1 and Ldb7, which were validated with a lipid dye recycling assay. Additionally, the essential phosphatidylinositol-4-kinase Pik1 was shown to have a role in recycling. We propose recycling is governed by Rpd3 at the transcriptional level via multiple downstream target genes.Konstantina AmoiradakiKate R. BuntingKatherine M. PaineJosephine E. AyreKaren HoggKamilla M. E. LaidlawChris MacDonaldMDPI AGarticleendocytosismembrane traffickingsurface membrane proteinshistone deacetylasetranscriptionBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12477, p 12477 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	endocytosis membrane trafficking surface membrane proteins histone deacetylase transcription Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	endocytosis membrane trafficking surface membrane proteins histone deacetylase transcription Biology (General) QH301-705.5 Chemistry QD1-999 Konstantina Amoiradaki Kate R. Bunting Katherine M. Paine Josephine E. Ayre Karen Hogg Kamilla M. E. Laidlaw Chris MacDonald The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
description	Intracellular trafficking pathways control residency and bioactivity of integral membrane proteins at the cell surface. Upon internalisation, surface cargo proteins can be delivered back to the plasma membrane via endosomal recycling pathways. Recycling is thought to be controlled at the metabolic and transcriptional level, but such mechanisms are not fully understood. In yeast, recycling of surface proteins can be triggered by cargo deubiquitination and a series of molecular factors have been implicated in this trafficking. In this study, we follow up on the observation that many subunits of the Rpd3 lysine deacetylase complex are required for recycling. We validate ten Rpd3-complex subunits in recycling using two distinct assays and developed tools to quantify both. Fluorescently labelled Rpd3 localises to the nucleus and complements recycling defects, which we hypothesised were mediated by modulated expression of Rpd3 target gene(s). Bioinformatics implicated 32 candidates that function downstream of Rpd3, which were over-expressed and assessed for capacity to suppress recycling defects of <i>rpd3</i>∆ cells. This effort yielded three hits: Sit4, Dit1 and Ldb7, which were validated with a lipid dye recycling assay. Additionally, the essential phosphatidylinositol-4-kinase Pik1 was shown to have a role in recycling. We propose recycling is governed by Rpd3 at the transcriptional level via multiple downstream target genes.
format	article
author	Konstantina Amoiradaki Kate R. Bunting Katherine M. Paine Josephine E. Ayre Karen Hogg Kamilla M. E. Laidlaw Chris MacDonald
author_facet	Konstantina Amoiradaki Kate R. Bunting Katherine M. Paine Josephine E. Ayre Karen Hogg Kamilla M. E. Laidlaw Chris MacDonald
author_sort	Konstantina Amoiradaki
title	The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
title_short	The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
title_full	The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
title_fullStr	The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
title_full_unstemmed	The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast
title_sort	rpd3-complex regulates expression of multiple cell surface recycling factors in yeast
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ea2cd7d020234a359aaee50b401a06dc
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The Rpd3-Complex Regulates Expression of Multiple Cell Surface Recycling Factors in Yeast

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