Changes of Nuclear Protein Kinase C Activity and Isotype Composition in PC12 Cell Proliferation and Differentiation

Borgatti, Paola; Mazzoni, M.P.; Carini, Cinzia; Neri, Luca M.; Marchisio, Marco; Bertolaso, Lucia; Previati, Maurizio; Zauli, Giorgio; Capitani, Silvano

doi:10.1006/excr.1996.0112

Cited by 58 publications

(47 citation statements)

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“…The supernatant from the first centrifugation was saved and used as cytoplasmic fraction. The purity of nuclear preparations was evaluated by Western blot analysis using a monoclonal antibody to b-tubulin, as previously described (Borgatti et al, 1996). The absence of immunoreactivity to the cytoskeletal protein in the isolated nuclear preparations confirmed that the isolation procedure yielded nuclei of high purity that were free of cytoplasmic contaminants (data not shown).…”

Section: Isolation Of Nucleimentioning

confidence: 75%

“…This was accomplished as previously reported, with minor changes (Borgatti et al, 1996). Briefly, cells (5 Â 10 6 ) were resuspended in 10 mM Tris-HCl, pH 7.8, 10 mM NaCl, 2 mM MgCl 2 , 10 mM b-mercaptoethanol, 0.5 mM phenylmethyl fluoride (PMSF), 10 mg/ml soybean trypsin inhibitor, 1 mg/ml of leupeptin and aprotinin, 1.0 mM Na 3 VO 4 , 20 nM okadaic acid.…”

Section: Isolation Of Nucleimentioning

confidence: 98%

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Threonine 308 phosphorylated form of akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin

Borgatti

Martelli

Tabellini³

et al. 2003

Journal Cellular Physiology

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We have examined the issue of whether or not in PC12 cells it may be observed a nerve growth factor (NGF) nuclear translocation of an active (phosphorylated) Akt. Western blot analysis with antibodies to either total or phosphorylated Akt showed a maximal nuclear translocation after 15 min of NGF stimulation. NGF increased rapidly and transiently the enzymatic activity of immunoprecipitable nuclear Akt and after 45 min the values returned to a level close to the basal one. Enzyme translocation was blocked by the selective phosphoinositide 3-kinase inhibitor, LY294002. Confocal microscopy of samples stained with antibody to Akt showed an evident increase in immunostaining intensity in the nuclear interior after NGF treatment. Treatment of cells with inhibitors of protein phosphatase PP2A, calyculin A, or okadaic acid, maintained the phosphorylation levels of nuclear Akt. Immunoprecipitation experiments revealed an association between Akt and PP2A that was maximal when nuclear Akt activity was decreased. Both total and active Akt associated with the nuclear matrix and, in particular, with the protein nucleolin, with which Akt co-immunoprecipitated. These findings strongly suggest that the intranuclear translocation of active Akt is an important step in the signaling pathways elicited by the neurotrophin NGF and that the intranuclear control of Akt is achieved through the action of PP2A.

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Section: Isolation Of Nucleimentioning

confidence: 75%

Section: Isolation Of Nucleimentioning

confidence: 98%

Threonine 308 phosphorylated form of akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin

Borgatti

Martelli

Tabellini³

et al. 2003

Journal Cellular Physiology

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“…It should be noted that, although insensitivity to direct activation by DAG has served to define atypical PKC isoforms, stimulation of PKC by PMA in various cell types including PC12 cells has been observed (64,65). Several studies have demonstrated that translocation of PKC to the nucleus is concurrent with activation (42,44,48,55). This facet of PKC activity is particularly relevant to VEGF mRNA regulation, as PKC-mediated activation of nuclear transcription factors such as Sp-1 has a critical role in VEGF gene transcription (66,67).…”

Section: Discussionmentioning

confidence: 99%

“…Based on their requirements for activation, three PKC classes are defined: conventional (␣, ␤I, ␤II, ␥), which are activated by Ca 2ϩ and diacylglycerol (DAG); novel (␦, ⑀, , ), which are Ca 2ϩ -independent; and atypical (, , ), which are activated independently of Ca 2ϩ or DAG. PC12 cells have been reported to express PKC isoforms ␣, ␤I, ␤II, ␦, ⑀, , , and (43)(44)(45). Two specific PKC isoforms, PKC and PKC⑀, have been demonstrated to regulate VEGF expression.…”

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confidence: 99%

Distinct Protein Kinase C Isoforms Mediate Regulation of Vascular Endothelial Growth Factor Expression by A2A Adenosine Receptor Activation and Phorbol Esters in Pheochromocytoma PC12 Cells

Gardner

Olah

2003

Journal of Biological Chemistry

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“…There are 12 different known PKC isozymes (Dempsey et al, 2000) and PC12 expresses at least seven of these PKC isozymes (Borgatti et al, 1996). twnt3 -4 cells were cultured in tetracycline-free medium to allow Wnt-1 expression.…”

Section: Discussionmentioning

confidence: 99%

Inhibition by Wnt-1 or Wnt-3a of nerve growth factor-induced differentiation of PC12 cells is reversed by bisindolylmaleimide-I but not by several other PKC inhibitors

Chou

Howard

2002

Oncogene

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Wnt-1 and Wnt-3a exhibit redundancy in neural crest development. We have found that they do not produce the same effects on PC12 cells, which were obtained from the adrenal medulla, a neural crest derivative. However, both Wnt-1 or Wnt-3a inhibit nerve growth factor (NGF)-induced neurite outgrowth. The inhibition is reversed by the protein kinase C (PKC) inhibitor, bisindolylmaleimide-I, but it did not reverse Wnt-1-induced activation of the canonical Wnt pathway. The Wnt-1 inhibitory effect was not reversed by several other PKC inhibitors, by phorbol ester-induced down-regulation of PKC, or by pertussis toxin, which is known to inhibit another Wnt signaling pathway, the Wnt/Ca 2+ pathway. We suggest that bisindolylmaleimide-I acts by affecting either a pathway downstream from Lef-1/Tcf in the canonical pathway or a Wnt signaling pathway other than the canonical pathway. In either case, the bisindolylmaleimide-I sensitivity of this pathway should aid in its identification.

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Changes of Nuclear Protein Kinase C Activity and Isotype Composition in PC12 Cell Proliferation and Differentiation

Cited by 58 publications

References 0 publications

Threonine 308 phosphorylated form of akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin

Threonine 308 phosphorylated form of akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin

Distinct Protein Kinase C Isoforms Mediate Regulation of Vascular Endothelial Growth Factor Expression by A2A Adenosine Receptor Activation and Phorbol Esters in Pheochromocytoma PC12 Cells

Inhibition by Wnt-1 or Wnt-3a of nerve growth factor-induced differentiation of PC12 cells is reversed by bisindolylmaleimide-I but not by several other PKC inhibitors

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