2. Academic Validation
  3. Identification of phosphorylation site on PARP1 mediating its cytosolic translocation in virus-infected HeLa cells

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

  • STAR Protoc. 2022 Nov 2;3(4):101808. doi: 10.1016/j.xpro.2022.101808.
Fei Wang 1 2 3,  Ming Tong Ma 1 2 3,  Junfang Xu 3,  Haipeng Liu 1 2 3 4
Affiliations

Affiliations

  • 1 Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, China.
  • 2 Department of Microbiology and Immunology, Tongji University School of Medicine, Shanghai 200072, China.
  • 3 Clinical Translation Research Center, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, China.
  • 4 Central Laboratory, Shanghai Pulmonary Hospital, School of Medicine, Tongji University School of Medicine, Shanghai 200433, China.
PMID: 36386864 DOI: 10.1016/j.xpro.2022.101808
Abstract

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).

Keywords

Cell biology; Cell separation/fractionation; Mass spectrometry; Microscopy; Molecular biology; Signal transduction.

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