Phosphoinositide-specific phospholipase C beta1 signal transduction in the nucleus

The nuclear inositol lipid cycle is a well known process, and nuclear phosphoinositide-specific phospholipase C beta1 (PLCbeta1) signalling activity has been extensively studied in the last decades. We now know that nuclear PLCbeta1 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 PLCbeta1 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 PLCbeta1 signaling. The described techniques range from nuclear purification to enzymatic activity and to molecular biology methods.