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PMID:17022104

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Citation

Bavelloni, A, Faenza, I, Cioffi, G, Piazzi, M, Parisi, D, Matic, I, Maraldi, NM and Cocco, L (2006) Proteomic-based analysis of nuclear signaling: PLCbeta1 affects the expression of the splicing factor SRp20 in Friend erythroleukemia cells. Proteomics 6:5725-34

Abstract

An extensive body of evidence links inositide-specific phospholipase C (PLC) to the nucleus and the main isoform located in the nucleus is PLCbeta(1). Constitutive overexpression of nuclear PLCbeta(1) has been previously shown to inhibit Friend erythroleukemia cells differentiation and to induce cell cycle progression targeting cyclin D3. The aim of this study was to identify new proteins regulated by PLCbeta(1) overexpression, given the role exerted by its signaling in the nucleus during cell growth and differentiation. To identify novel downstream effectors of nuclear PLCbeta(1)-dependent signaling in Friend erythroleukemia cells, we performed the high-resolution 2-DE-based proteomic analysis. Using a proteomic approach we found that SRp20, a member of the highly conserved SR family of splicing regulators, was down-regulated in cells overexpressing nuclear PLCbeta(1) as compared with wild-type cells. Reduction in SRp20 was confirmed by 2-D Western blotting. Moreover, we have shown that nuclear PLCbeta(1) is bound to the SRp20 splicing factor. Indeed, by immunoprecipitation and subcellular fractioning, we have demonstrated that endogenous PLCbeta(1) and SRp20 physically interact in the nucleus. Here we show the existence of a PLCbeta(1)-specific target, the splicing factor SRp20, whose expression is specifically down-regulated by the nuclear signaling evoked by PLCbeta(1).

Links

PubMed Online version:10.1002/pmic.200600318

Keywords

Animals; Blotting, Western; Cell Nucleus/metabolism; Cells, Cultured; Down-Regulation; Electrophoresis, Gel, Two-Dimensional; Fluorescein-5-isothiocyanate; Fluorescent Antibody Technique, Direct; Fluorescent Dyes; Gene Expression Regulation, Neoplastic; Isoelectric Focusing; Isoenzymes/genetics; Isoenzymes/metabolism; Leukemia, Erythroblastic, Acute/metabolism; Leukemia, Erythroblastic, Acute/pathology; Mice; Microscopy, Fluorescence; Peptide Mapping; Phospholipase C beta; Precipitin Tests; Proteomics/methods; RNA-Binding Proteins/metabolism; RNA-Binding Proteins/physiology; Signal Transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subcellular Fractions/metabolism; Type C Phospholipases/genetics; Type C Phospholipases/metabolism

Significance

Annotations

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

MOUSE:SRSF3

located_in

GO:0016607: nuclear speck

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:SRSF3

involved_in

GO:0008286: insulin receptor signaling pathway

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

MOUSE:SRSF3

enables

GO:0043274: phospholipase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9Z1B3

F

Seeded From UniProt

complete

MOUSE:PLCB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P84104

F

Seeded From UniProt

complete

MOUSE:PLCB1

located_in

GO:0016607: nuclear speck

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:PLCB1

involved_in

GO:0045892: negative regulation of transcription, DNA-templated

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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