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Phosphoinositide-Specific Phospholipase C β1 Signal Transduction in the Nucleus

  • Roberta Fiume
  • Gabriella Teti
  • Irene Faenza
  • Lucio Cocco
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 645)

Abstract

The nuclear inositol lipid cycle is a well known process, and nuclear phosphoinositide-specific phospholipase C β1 (PLCβ1) signalling activity has been extensively studied in the last decades. We now know that nuclear PLCβ1 is a key player in the control of cell cycle progression; in fact it appears to be involved in the cyclin-mediated regulation of the physiological machinery. Indeed, the recent discovery of a possible involvement of the interstitial deletion of PLCβ1 gene in the progression of myelodysplastic syndrome (MDS) to acute myeloid leukemia in humans (AML) strengthens this contention.

Albeit several papers have reported the techniques used for the study of inositide-dependent signaling in the nucleus, we describe here step by step protocols, which can be followed for the preparation of highly purified nuclei and the subsequent analysis of nuclear PLCβ1 signaling. The described techniques range from nuclear purification to enzymatic activity and to molecular biology methods.

Key words:

Phospholipase C β1 Nuclear signaling PLC activity assay Nuclei purification Transmission electron microscopy PLCβ1a and 1b alternative splicing 

Notes

Acknowledgments

The authors own work was supported by grants from Italian MIUR-FIRB (HumanProteome Net), MIUR-COFIN, and the Carisbo Foundation.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Roberta Fiume
    • 1
  • Gabriella Teti
    • 2
  • Irene Faenza
    • 1
  • Lucio Cocco
    • 1
  1. 1.Cellular Signaling Laboratory, Department of Anatomical SciencesUniversity of BolognaBolognaItaly
  2. 2.Department of Anatomical SciencesUniversity of BolognaBolognaItaly

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