Identification of phosphoprotein NP33 as a nucleus-associated ribosomal S6 protein and its phosphorylation in hematopoietic cells

Donald E Macfarlane; David Gailani

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Identification of phosphoprotein NP33 as a nucleus-associated ribosomal S6 protein and its phosphorylation in hematopoietic cells

Journal article

Peer reviewed

Identification of phosphoprotein NP33 as a nucleus-associated ribosomal S6 protein and its phosphorylation in hematopoietic cells

Donald E Macfarlane and David Gailani

Cancer research (Chicago, Ill.), Vol.50(10), pp.2895-2900

05/15/1990

PMID: 2334893

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Abstract

Exposure of HL-60 promyelocytes to the phorbol ester 12-O-tetradecanoylphorbol-13-acetate increased incorporation of 32P into a Mr approximately 33,000 protein (NP33) found in the nuclear matrices prepared by treating cells with Triton X-100, nucleases, and 2 M NaCl (D. E. Macfarlane, J. Biol. Chem., 261: 6947-6953, 1986). We now report that 12-O-tetradecanoylphorbol-13-acetate causes phosphorylation of NP33 in U937, K562, HEL, Molt-3, and Raji cell lines, all of which are rapidly proliferating cells of hematopoietic origin. 12-O-Tetradecanoylphorbol-13-acetate caused a lesser degree of NP33 phosphorylation in peripheral blood lymphocytes and monocytes and none in granulocytes or platelets. The incorporation of 32P into NP33 was complete in about 10 min, and it was prevented or reversed by staurosporin, indicating that NP33 is continuously phosphorylated and dephosphorylated. NP33 was purified to homogeneity from Triton X-100-washed nuclei or whole cells by extraction with H2SO4, acetone precipitation, and preparative two-dimensional gel electrophoresis. The amino-terminal amino acid sequence of NP33 appears to be the same as that of ribosomal S6 protein. NP33 appears to be S6 protein copurifying with the nuclear matrix.

Phosphorylation

Ribosomal Proteins - analysis

Amino Acid Sequence

Cell Line

Alkaloids - pharmacology

Nuclear Proteins - analysis

Tetradecanoylphorbol Acetate - pharmacology

Staurosporine

Humans

Molecular Sequence Data

Electrophoresis, Gel, Two-Dimensional

Nuclear Proteins - metabolism

Hematopoietic Stem Cells - metabolism

Phosphoproteins - metabolism

Ribosomal Proteins - metabolism

Protein Kinase Inhibitors

Phosphoproteins - analysis

Ribosomal Protein S6

Details

Title: Subtitle: Identification of phosphoprotein NP33 as a nucleus-associated ribosomal S6 protein and its phosphorylation in hematopoietic cells
Creators: Donald E Macfarlane - Department of Medicine, Veterans Administration Medical Center, Iowa City, Iowa
David Gailani
Resource Type: Journal article
Publication Details: Cancer research (Chicago, Ill.), Vol.50(10), pp.2895-2900
PMID: 2334893
ISSN: 0008-5472
eISSN: 1538-7445
Grant note: T32 HLO 7344 / NHLBI NIH HHS
Language: English
Date published: 05/15/1990
Academic Unit: Hematology, Oncology, and Blood & Marrow Transplantation; Internal Medicine
Record Identifier: 9984094767502771

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