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PMID:11082283

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Citation

Roymans, D, Willems, R, Vissenberg, K, De Jonghe, C, Grobben, B, Claes, P, Lascu, I, Van Bockstaele, D, Verbelen, JP, Van Broeckhoven, C and Slegers, H (2000) Nucleoside diphosphate kinase beta (Nm23-R1/NDPKbeta) is associated with intermediate filaments and becomes upregulated upon cAMP-induced differentiation of rat C6 glioma. Exp. Cell Res. 261:127-38

Abstract

Nucleoside diphosphate kinases (Nm23/NDPK) are enzymes functional in cell proliferation, differentiation, development, tumor progression, and metastasis. Nevertheless, no consensus exists about the molecular mechanism by which Nm23/NDPK isoforms exert their role in these processes. We investigated the expression of the rat Nm23-R1/NDPKbeta and Nm23-R2/NDPKalpha isoforms, homologues of the human Nm23-H1/NDPK A and Nm23-H2/NDPK B proteins, respectively, upon cAMP-induced differentiation of rat C6 glioma cells and demonstrated a differential interaction with intermediate filaments. Semiquantitative RT-PCR, immunoblotting, and flow cytometry showed a constitutive expression of both Nm23 isoforms. After induction of differentiation in C6 cells with cAMP analogs or isoproterenol, a dose-dependent 2- and 2.5-fold upregulation of the Nm23-R1 mRNA and protein, respectively, was observed. In contrast, the expression of Nm23-R2 remained unchanged. Localization of both isoforms with confocal laser scanning microscopy demonstrated a punctate reticular staining pattern for both Nm23 isoforms in the cytosol and processes of the cells which was particularly intense in the perinuclear region. In addition, while Nm23-R2 was colocalized and coimmunoprecipitated with vimentin in nondifferentiated cells, both isoforms were associated with GFAP in differentiated cells. The significance of these findings in relation to a possible function of Nm23 isoforms in cell proliferation, differentiation, and tumor-associated mechanisms is discussed.

Links

PubMed Online version:10.1006/excr.2000.5037

Keywords

8-Bromo Cyclic Adenosine Monophosphate/pharmacology; Animals; Bucladesine/pharmacology; Cell Differentiation/physiology; Cyclic AMP/physiology; Cytosol/enzymology; Gene Expression Regulation, Enzymologic/drug effects; Glioma; Humans; Intermediate Filaments/physiology; Isoenzymes/genetics; Isoenzymes/metabolism; Isoproterenol/pharmacology; Kinetics; Monomeric GTP-Binding Proteins/genetics; Monomeric GTP-Binding Proteins/metabolism; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase/genetics; Nucleoside-Diphosphate Kinase/metabolism; Protein Biosynthesis; Rats; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors/genetics; Transcription Factors/metabolism; Transcription, Genetic; Tumor Cells, Cultured

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:NDKB

enables

GO:0019215: intermediate filament binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:621646

F

Seeded From UniProt

complete

RAT:NDKB

enables

GO:0019215: intermediate filament binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:2679

F

Seeded From UniProt

complete

RAT:NDKB

located_in

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:NDKB

colocalizes_with

GO:0005882: intermediate filament

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:VIME

enables

GO:0019900: kinase binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:619877

F

Seeded From UniProt

complete

RAT:GFAP

enables

GO:0019900: kinase binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:70497

F

Seeded From UniProt

complete

RAT:GFAP

enables

GO:0019900: kinase binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:619877

F

Seeded From UniProt

complete

RAT:NDKA

located_in

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:NDKA

colocalizes_with

GO:0005882: intermediate filament

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:NDKA

enables

GO:0019215: intermediate filament binding

ECO:0000353: physical interaction evidence used in manual assertion

RGD:2679

F

Seeded From UniProt

complete

RAT:NDKA

involved_in

GO:0035690: cellular response to drug

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:NDKA

involved_in

GO:0042493: response to drug

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:NDKA

part_of

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:NDKA

involved_in

GO:0051591: response to cAMP

ECO:0000270: expression pattern evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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