Effects of adenylyl cyclase and protein kinase A inhibition on signaling enzymes in natural killer cells: comparison to tributyltin.
Natural killer (NK) cells are lymphocytes capable of destroying tumor cells and virally-infected cells without prior sensitization. In a previous study, we found that inhibition of adenylyl cyclase (AC) or cAMP-dependent protein kinase (PKA) decreased the ability of NK cells to destroy tumor cells. We also found that the environmental contaminant tributyltin (TBT), at concentrations of 300-500 nM, decreased tumor-cell lysis by NK cells, as well as their intracellular levels of cAMP. This suggested that the decreases in cAMP associated with TBT (300-500 nM) may, in part, be responsible for loss of cytotoxic function. Here, we investigated the effects of inhibition of AC or PKA on enzymes that are required in the NK tumorolytic process and compared them to those of TBT exposure. The enzymes studied were: the protein tyrosine kinase (PTK), syk; phospholipase C gamma1 (PLCgamma1); and the mitogen activated protein kinase (MAPK), p44/42. Exposure of NK cells to the AC inhibitor 2',5'-dideoxyadenosine (DDA) significantly increased the total level of PLCgamma1 by 67% after 60 min and the level of p44/42 by about 30%. Exposure to the PKA inhibitor H-89 significantly increased the levels of the phosphorylated (activated) p44/42 (90%) after 60 min. Exposure to TBT increased the levels of PLCgamma1 by about 50%. Previously, we found that exposure to TBT increased the phosphorylation of p44/42 within 5 min. These results indicate that AC inhibition caused alterations of the levels of key enzymes, while decreased PKA activity caused an increase in p44/42 activation. They also suggest that the effects of decreased levels of cAMP on these key cytotoxic signaling proteins may overlap, to a very limited extent, with those of TBT.
Whalen
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MM
Department of Chemistry, Tennessee State University, Nashville, TN 37209, USA. mwhalen@tnstate.edu
Odman-Ghazi
S O
SO
D000067956:Adenylyl Cyclase Inhibitors; D000998:Antiviral Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016048:Dideoxyadenosine / Q000494:pharmacology*; D006801:Humans; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000201:enzymology; D010766:Phosphorylation / Q000187:drug effects; D014220:Trialkyltin Compounds / Q000494:pharmacology*
D000067956:Adenylyl Cyclase Inhibitors; D000998:Antiviral Agents; D014220:Trialkyltin Compounds; D016048:Dideoxyadenosine; C011559:tributyltin; D017868:Cyclic AMP-Dependent Protein Kinases