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Multiphosphorylation and cellular localization of poly(rC) binding protein 1 during mitosis in hela cell

Authors
  • Ma, Xiang1
  • Li, Huicai2
  • Cao, Xue2
  • Liu, Zhu1
  • He, Dacheng2
  • 1 Hainan University, Key Laboratory of Tropical Biological Resources of Ministry of Education, School of Life and Pharmaceutical Sciences, Haikou, 570228, China , Haikou (China)
  • 2 Beijing Normal University, Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing, 100875, China , Beijing (China)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
May 10, 2019
Volume
41
Issue
6-7
Pages
711–717
Identifiers
DOI: 10.1007/s10529-019-02679-1
Source
Springer Nature
Keywords
License
Yellow

Abstract

ObjectiveTo monitor the phosphorylation modifications and cellular localization of poly(rC)-binding protein-1 (PCBP1) during the cell cycle progression of Hela cells.ResultHela cells highly synchronized at five different phases from interphase to mitosis were obtained. Using mitotic phosphoprotein-specific monoclonal antibody MPM-2, the exclusive occurrences of multiphosphorylation statuses of PCBP1 in mitosis were confirmed by a series of spots with increasing acidic pI (isoelectric point) in two rounds of 2D western blotting on the same membrane, and a visible molecular mass shift that can be eliminated by the treatment with λ phosphatase in 1D western blotting. Immnuofluorescence revealed the localization shift of PCBP1 during cell cycle, with accumulations in nucleus as a patch pattern in interphase, and a dispersive distribution without the area of the condensed chromosomes during mitosis.ConclusionsThese observations of mitosis-specific multiphosphorylations and localization shifts of PCBP1 suggest that the versatile PCBP1 was regulatable in a phosphorylation modification- and temporospatial-dependent manner in mitotic regulatory networks.

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