Cargando…

Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells

Poly (ADP-ribose) polymerase 1 (PARP1) localization is controlled by its phosphorylation state. Here, we describe a protocol to monitor PARP1 subcellular localization in HSV-1-infected HeLa cells using immunofluorescence microscopy and cytoplasmic/nuclear fractionation. We detail steps to identify p...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Fei, Ma, Ming Tong, Xu, Junfang, Liu, Haipeng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9640338/
https://www.ncbi.nlm.nih.gov/pubmed/36386864
http://dx.doi.org/10.1016/j.xpro.2022.101808
_version_ 1784825827844685824
author Wang, Fei
Ma, Ming Tong
Xu, Junfang
Liu, Haipeng
author_facet Wang, Fei
Ma, Ming Tong
Xu, Junfang
Liu, Haipeng
author_sort Wang, Fei
collection PubMed
description Poly (ADP-ribose) polymerase 1 (PARP1) localization is controlled by its phosphorylation state. Here, we describe a protocol to monitor PARP1 subcellular localization in HSV-1-infected HeLa cells using immunofluorescence microscopy and cytoplasmic/nuclear fractionation. We detail steps to identify phosphorylation sites on PARP1 using conserved motif analysis and mass spectrometry. This protocol can be applied to the study of other protein phosphorylation events in other cell types. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022).
format Online
Article
Text
id pubmed-9640338
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-96403382022-11-15 Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells Wang, Fei Ma, Ming Tong Xu, Junfang Liu, Haipeng STAR Protoc Protocol Poly (ADP-ribose) polymerase 1 (PARP1) localization is controlled by its phosphorylation state. Here, we describe a protocol to monitor PARP1 subcellular localization in HSV-1-infected HeLa cells using immunofluorescence microscopy and cytoplasmic/nuclear fractionation. We detail steps to identify phosphorylation sites on PARP1 using conserved motif analysis and mass spectrometry. This protocol can be applied to the study of other protein phosphorylation events in other cell types. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022). Elsevier 2022-11-02 /pmc/articles/PMC9640338/ /pubmed/36386864 http://dx.doi.org/10.1016/j.xpro.2022.101808 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Wang, Fei
Ma, Ming Tong
Xu, Junfang
Liu, Haipeng
Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title_full Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title_fullStr Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title_full_unstemmed Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title_short Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells
title_sort identification of phosphorylation site on parp1 mediating its cytosolic translocation in virus-infected hela cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9640338/
https://www.ncbi.nlm.nih.gov/pubmed/36386864
http://dx.doi.org/10.1016/j.xpro.2022.101808
work_keys_str_mv AT wangfei identificationofphosphorylationsiteonparp1mediatingitscytosolictranslocationinvirusinfectedhelacells
AT mamingtong identificationofphosphorylationsiteonparp1mediatingitscytosolictranslocationinvirusinfectedhelacells
AT xujunfang identificationofphosphorylationsiteonparp1mediatingitscytosolictranslocationinvirusinfectedhelacells
AT liuhaipeng identificationofphosphorylationsiteonparp1mediatingitscytosolictranslocationinvirusinfectedhelacells