Phosphoinositide-Specific Phospholipase C β1 Signal Transduction in the Nucleus
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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
The authors own work was supported by grants from Italian MIUR-FIRB (HumanProteome Net), MIUR-COFIN, and the Carisbo Foundation.
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