Manipulation of PKC isozymes by RNA interference and inducible expression of PKC constructs.
Protein kinase C (PKC), a family of serine/threonine kinases, plays an important role in apoptosis. Several members of the PKC family act as substrates for caspases. In addition, PKCs can also regulate caspase activation and cell death by apoptosis. The cleavage of PKCs separates the regulatory domain from the catalytic domain. The full-length, the catalytic domain, and the regulatory domain of PKC family members may have distinct function in apoptosis. Delineating the role of protein kinase C (PKC) isozymes in apoptosis has been challenging because of the lack of selective inhibitors of PKC isozymes and difficulty in generating stable cell lines expressing pro-apoptotic PKC isozymes. In this chapter, we describe the use of RNA interference (siRNA) technology and tetracycline-inducible expression of PKC isozymes to study their function in apoptosis.
Basu
Alakananda
A
Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, Texas, USA.
Persaud
Shalini D
SD
Sivaprasad
Usha
U
D017209:Apoptosis / Q000502:physiology*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D007527:Isoenzymes / Q000235:genetics; D011493:Protein Kinase C / Q000096:biosynthesis / Q000235:genetics*; D034622:RNA Interference / Q000502:physiology*; D034741:RNA, Small Interfering / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology; D015534:Trans-Activators / Q000502:physiology; D014162:Transfection / Q000379:methods
D007527:Isoenzymes; D034741:RNA, Small Interfering; D015534:Trans-Activators; C415845:VP16-E protein, Equine herpesvirus 1; D011493:Protein Kinase C; D013752:Tetracycline