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Role of Regulatory Subunits and Protein Kinase Inhibitor (PKI) in Determining Nuclear Localization and Activity of the Catalytic Subunit of Protein Kinase A -- Wiley et al. 274 (10): 6381 -- Journal of Biological Chemistry
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Articles by Wiley, J. C.
Articles by McKnight, G. S.
J Biol Chem, Vol. 274, Issue 10, 6381-6387, March 5, 1999
Role of Regulatory Subunits and Protein Kinase Inhibitor
(PKI) in Determining Nuclear Localization and Activity of the
Catalytic Subunit of Protein Kinase A
Jesse C.
Wiley,
Lauren A.
Wailes,
Rejean L.
Idzerda, and
G.
Stanley
McKnight
From the Department of Pharmacology, University of
Washington, Seattle, Washington 98195-7750
Regulation of protein kinase A by subcellular
localization may be critical to target catalytic subunits to specific
substrates. We employed epitope-tagged catalytic subunit to correlate
subcellular localization and gene-inducing activity in the presence of
regulatory subunit or protein kinase inhibitor (PKI). Transiently
expressed catalytic subunit distributed throughout the cell and induced gene expression. Co-expression of regulatory subunit or PKI blocked gene induction and prevented nuclear accumulation. A mutant PKI lacking
the nuclear export signal blocked gene induction but not nuclear
accumulation, demonstrating that nuclear export is not essential to
inhibit gene induction. When the catalytic subunit was targeted to the
nucleus with a nuclear localization signal, it was not sequestered in
the cytoplasm by regulatory subunit, although its activity was
completely inhibited. PKI redistributed the nuclear catalytic subunit
to the cytoplasm and blocked gene induction, demonstrating that the
nuclear export signal of PKI can override a strong nuclear localization
signal. With increasing PKI, the export process appeared to saturate,
resulting in the return of catalytic subunit to the nucleus. These
results demonstrate that both the regulatory subunit and PKI are able
to completely inhibit the gene-inducing activity of the catalytic
subunit even when the catalytic subunit is forced to concentrate in the
nuclear compartment.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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