Cargando…

Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy

Background: The ARF tumour suppressor plays a well-established role as a tumour suppressor, halting cell growth by both p53-dependent and independent pathways in several cellular stress response circuits. However, data collected in recent years challenged the traditional role of this protein as a tu...

Descripción completa

Detalles Bibliográficos
Autores principales: Fontana, Rosa, Guidone, Daniela, Angrisano, Tiziana, Calabrò, Viola, Pollice, Alessandra, La Mantia, Girolama, Vivo, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8773949/
https://www.ncbi.nlm.nih.gov/pubmed/35053274
http://dx.doi.org/10.3390/biom12010126
_version_ 1784636223637159936
author Fontana, Rosa
Guidone, Daniela
Angrisano, Tiziana
Calabrò, Viola
Pollice, Alessandra
La Mantia, Girolama
Vivo, Maria
author_facet Fontana, Rosa
Guidone, Daniela
Angrisano, Tiziana
Calabrò, Viola
Pollice, Alessandra
La Mantia, Girolama
Vivo, Maria
author_sort Fontana, Rosa
collection PubMed
description Background: The ARF tumour suppressor plays a well-established role as a tumour suppressor, halting cell growth by both p53-dependent and independent pathways in several cellular stress response circuits. However, data collected in recent years challenged the traditional role of this protein as a tumour suppressor. Cancer cells expressing high ARF levels showed that its expression, far from being dispensable, is required to guarantee tumour cell survival. In particular, ARF can promote autophagy, a self-digestion pathway that helps cells cope with stressful growth conditions arising during both physiological and pathological processes. Methods: We previously showed that ARF is regulated through the activation of the protein kinase C (PKC)-dependent pathway and that an ARF phospho-mimetic mutant on the threonine residue 8, ARF-T8D, sustains cell proliferation in HeLa cells. We now explored the role of ARF phosphorylation in both basal and starvation-induced autophagy by analysing autophagic flux in cells transfected with either WT and ARF phosphorylation mutants by immunoblot and immunofluorescence. Results: Here, we show that endogenous ARF expression in HeLa cells is required for starvation-induced autophagy. Further, we provide evidence that the hyper-expression of ARF-T8D appears to inhibit autophagy in both HeLa and lung cancer cells H1299. This effect is due to the cells’ inability to elicit autophagosomes formation upon T8D expression. Conclusions: Our results lead to the hypothesis that ARF phosphorylation could be a mechanism through which the protein promotes or counteracts autophagy. Several observations underline how autophagy could serve a dual role in cancer progression, either protecting healthy cells from damage or aiding cancerous cells to survive. Our results indicate that ARF phosphorylation controls protein’s ability to promote or counteract autophagy, providing evidence of the dual role played by ARF in cancer progression.
format Online
Article
Text
id pubmed-8773949
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87739492022-01-21 Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy Fontana, Rosa Guidone, Daniela Angrisano, Tiziana Calabrò, Viola Pollice, Alessandra La Mantia, Girolama Vivo, Maria Biomolecules Communication Background: The ARF tumour suppressor plays a well-established role as a tumour suppressor, halting cell growth by both p53-dependent and independent pathways in several cellular stress response circuits. However, data collected in recent years challenged the traditional role of this protein as a tumour suppressor. Cancer cells expressing high ARF levels showed that its expression, far from being dispensable, is required to guarantee tumour cell survival. In particular, ARF can promote autophagy, a self-digestion pathway that helps cells cope with stressful growth conditions arising during both physiological and pathological processes. Methods: We previously showed that ARF is regulated through the activation of the protein kinase C (PKC)-dependent pathway and that an ARF phospho-mimetic mutant on the threonine residue 8, ARF-T8D, sustains cell proliferation in HeLa cells. We now explored the role of ARF phosphorylation in both basal and starvation-induced autophagy by analysing autophagic flux in cells transfected with either WT and ARF phosphorylation mutants by immunoblot and immunofluorescence. Results: Here, we show that endogenous ARF expression in HeLa cells is required for starvation-induced autophagy. Further, we provide evidence that the hyper-expression of ARF-T8D appears to inhibit autophagy in both HeLa and lung cancer cells H1299. This effect is due to the cells’ inability to elicit autophagosomes formation upon T8D expression. Conclusions: Our results lead to the hypothesis that ARF phosphorylation could be a mechanism through which the protein promotes or counteracts autophagy. Several observations underline how autophagy could serve a dual role in cancer progression, either protecting healthy cells from damage or aiding cancerous cells to survive. Our results indicate that ARF phosphorylation controls protein’s ability to promote or counteract autophagy, providing evidence of the dual role played by ARF in cancer progression. MDPI 2022-01-13 /pmc/articles/PMC8773949/ /pubmed/35053274 http://dx.doi.org/10.3390/biom12010126 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Fontana, Rosa
Guidone, Daniela
Angrisano, Tiziana
Calabrò, Viola
Pollice, Alessandra
La Mantia, Girolama
Vivo, Maria
Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title_full Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title_fullStr Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title_full_unstemmed Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title_short Mutation of the Conserved Threonine 8 within the Human ARF Tumour Suppressor Protein Regulates Autophagy
title_sort mutation of the conserved threonine 8 within the human arf tumour suppressor protein regulates autophagy
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8773949/
https://www.ncbi.nlm.nih.gov/pubmed/35053274
http://dx.doi.org/10.3390/biom12010126
work_keys_str_mv AT fontanarosa mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT guidonedaniela mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT angrisanotiziana mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT calabroviola mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT pollicealessandra mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT lamantiagirolama mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy
AT vivomaria mutationoftheconservedthreonine8withinthehumanarftumoursuppressorproteinregulatesautophagy