Interaction of protein phosphatase 1 delta with nucleolin in human osteoblastic cells

Hiroyuki Morimoto, Hirohiko Okamura, Tatsuji Haneji

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

We examined the expression and cytolocalization of the protein phosphatase type 1 delta (PP1δ) isoform and nucleolin in human osteoblastic MG63 and Saos-2 cells. Cellular fractionation of MG63 cells was done and protein was prepared from each fraction. Anti-nucleolin antibody interacted with the 100- and 95-kD proteins present in the whole-cell lysate. The 100-kD protein was detected in nuclear and nucleolar fractions. The 95-kD protein was detected in cytosolic and nucleoplasmic fractions. PP1δ and nucleolin were colocalized in the nucleolus in MG63 and Saos-2 cells revealed by an immunofluorescence method. PP1δ and nucleolin were also co-immunoprecipitated with anti-nucleolin and antiPP1δ antibodies. In the actinomycin D-treated cells, the subcellular localization of PP1δ and nucleolin was changed. Expression of PP1δ was upregulated with actinomycin D treatment. The level of 100-kD protein did not change in the actinomycin D-treated cells. However, the level of the 95-kD band increased with actinomycin D treatment. These results indicate that PP1δ was associated with nucleolin in the nucleolus of MG63 and Saos-2 cells and that nucleolin is a possible candidate substrate for PP1δ.

Original languageEnglish
Pages (from-to)1187-1193
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume50
Issue number9
Publication statusPublished - Sep 2002
Externally publishedYes

Fingerprint

Protein Phosphatase 1
Dactinomycin
Proteins
Cell Fractionation
nucleolin
Fluorescent Antibody Technique
Anti-Idiotypic Antibodies
Protein Isoforms
Antibodies
Therapeutics

Keywords

  • Nucleolin
  • Osteoblast
  • Protein phosphatase

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Interaction of protein phosphatase 1 delta with nucleolin in human osteoblastic cells. / Morimoto, Hiroyuki; Okamura, Hirohiko; Haneji, Tatsuji.

In: Journal of Histochemistry and Cytochemistry, Vol. 50, No. 9, 09.2002, p. 1187-1193.

Research output: Contribution to journalArticle

@article{25e18c9ce13849c2bbaf5ae92e63afa8,
title = "Interaction of protein phosphatase 1 delta with nucleolin in human osteoblastic cells",
abstract = "We examined the expression and cytolocalization of the protein phosphatase type 1 delta (PP1δ) isoform and nucleolin in human osteoblastic MG63 and Saos-2 cells. Cellular fractionation of MG63 cells was done and protein was prepared from each fraction. Anti-nucleolin antibody interacted with the 100- and 95-kD proteins present in the whole-cell lysate. The 100-kD protein was detected in nuclear and nucleolar fractions. The 95-kD protein was detected in cytosolic and nucleoplasmic fractions. PP1δ and nucleolin were colocalized in the nucleolus in MG63 and Saos-2 cells revealed by an immunofluorescence method. PP1δ and nucleolin were also co-immunoprecipitated with anti-nucleolin and antiPP1δ antibodies. In the actinomycin D-treated cells, the subcellular localization of PP1δ and nucleolin was changed. Expression of PP1δ was upregulated with actinomycin D treatment. The level of 100-kD protein did not change in the actinomycin D-treated cells. However, the level of the 95-kD band increased with actinomycin D treatment. These results indicate that PP1δ was associated with nucleolin in the nucleolus of MG63 and Saos-2 cells and that nucleolin is a possible candidate substrate for PP1δ.",
keywords = "Nucleolin, Osteoblast, Protein phosphatase",
author = "Hiroyuki Morimoto and Hirohiko Okamura and Tatsuji Haneji",
year = "2002",
month = "9",
language = "English",
volume = "50",
pages = "1187--1193",
journal = "Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
publisher = "Histochemical Society Inc.",
number = "9",

}

TY - JOUR

T1 - Interaction of protein phosphatase 1 delta with nucleolin in human osteoblastic cells

AU - Morimoto, Hiroyuki

AU - Okamura, Hirohiko

AU - Haneji, Tatsuji

PY - 2002/9

Y1 - 2002/9

N2 - We examined the expression and cytolocalization of the protein phosphatase type 1 delta (PP1δ) isoform and nucleolin in human osteoblastic MG63 and Saos-2 cells. Cellular fractionation of MG63 cells was done and protein was prepared from each fraction. Anti-nucleolin antibody interacted with the 100- and 95-kD proteins present in the whole-cell lysate. The 100-kD protein was detected in nuclear and nucleolar fractions. The 95-kD protein was detected in cytosolic and nucleoplasmic fractions. PP1δ and nucleolin were colocalized in the nucleolus in MG63 and Saos-2 cells revealed by an immunofluorescence method. PP1δ and nucleolin were also co-immunoprecipitated with anti-nucleolin and antiPP1δ antibodies. In the actinomycin D-treated cells, the subcellular localization of PP1δ and nucleolin was changed. Expression of PP1δ was upregulated with actinomycin D treatment. The level of 100-kD protein did not change in the actinomycin D-treated cells. However, the level of the 95-kD band increased with actinomycin D treatment. These results indicate that PP1δ was associated with nucleolin in the nucleolus of MG63 and Saos-2 cells and that nucleolin is a possible candidate substrate for PP1δ.

AB - We examined the expression and cytolocalization of the protein phosphatase type 1 delta (PP1δ) isoform and nucleolin in human osteoblastic MG63 and Saos-2 cells. Cellular fractionation of MG63 cells was done and protein was prepared from each fraction. Anti-nucleolin antibody interacted with the 100- and 95-kD proteins present in the whole-cell lysate. The 100-kD protein was detected in nuclear and nucleolar fractions. The 95-kD protein was detected in cytosolic and nucleoplasmic fractions. PP1δ and nucleolin were colocalized in the nucleolus in MG63 and Saos-2 cells revealed by an immunofluorescence method. PP1δ and nucleolin were also co-immunoprecipitated with anti-nucleolin and antiPP1δ antibodies. In the actinomycin D-treated cells, the subcellular localization of PP1δ and nucleolin was changed. Expression of PP1δ was upregulated with actinomycin D treatment. The level of 100-kD protein did not change in the actinomycin D-treated cells. However, the level of the 95-kD band increased with actinomycin D treatment. These results indicate that PP1δ was associated with nucleolin in the nucleolus of MG63 and Saos-2 cells and that nucleolin is a possible candidate substrate for PP1δ.

KW - Nucleolin

KW - Osteoblast

KW - Protein phosphatase

UR - http://www.scopus.com/inward/record.url?scp=0036733594&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036733594&partnerID=8YFLogxK

M3 - Article

VL - 50

SP - 1187

EP - 1193

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

SN - 0022-1554

IS - 9

ER -