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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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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 |
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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 |
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