PIN: a novel protein involved in IFN-gamma accumulation of NOS-1 in neurons.
In this study we investigate the role of the protein inhibitor of NOS-1 (PIN) in the interferon-gamma (IFN-gamma)-mediated posttranscriptional accumulation of nitric oxide synthase-1 (NOS-1) and the anti-vesicular stomatitis virus response in neuronal cells. IFN-gamma-induced enhancement of NOS-1 activity is crucial for its antiviral activity in the central nervous system. IFN-gamma treatment of neuronal cells results in an increase of total NOS-1 and decrease of total PIN proteins without alteration in their respective mRNA levels. PIN/NOS-1 complexes decreased after IFN-gamma treatment. Transfection of cells with small interfering RNA (siRNA) for PIN results in a higher constitutive activity of NOS-1 and inhibition of viral replication. IFN-gamma treatment did not change the amount of NOS-1 detectable by Western blot, when PIN is diminished by RNAi treatment. Overexpression of PIN results in lower constitutive NOS-1 expression and activity, and diminishes activation of NOS-1 by IFN-gamma. Our findings indicate that in neurons, IFN-gamma upregulates NOS-1 through proteasomal degradation of PIN.
Yang
Jingjun
J
Department of Biology, New York University, New York, New York 10003-6688, USA.
Dennison
Natalie Nicole
NN
Reiss
Carol Shoshkes
CS
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D056445:Cytoplasmic Dyneins; D017931:DNA Primers / Q000737:chemistry; D004398:Dyneins / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology*; D051379:Mice; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D017471:Receptors, Interferon / Q000378:metabolism; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D009569:Nitric Oxide; D007371:Interferon-gamma; D052248:Nitric Oxide Synthase Type I; C486069:Dynll1 protein, rat; D056445:Cytoplasmic Dyneins; D004398:Dyneins