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Journal of Neuroscience, Vol 13, 5242-5250, Copyright © 1993 by Society for Neuroscience
PKA-dependent regulation of mKv1.1, a mouse Shaker-like potassium channel gene, when stably expressed in CHO cells
MM Bosma, ML Allen, TM Martin and BL Tempel
Geriatric Research Education and Clinical Center, VA Medical Center, Seattle, Washington 98108.
Potassium (K) channels are important regulators of cellular physiology and
can themselves be modulated by phosphorylation. We have investigated the
potential protein kinase A (PKA) regulation of mKv1.1, a mouse Shaker-like
K channel gene, when it is expressed in stably transfected Chinese hamster
ovary (CHO) cell lines. Whole-cell patch- clamp records show that
expression of mKv1.1 gives rise to a rapidly activating, sustained K+
current, referred to classically as a delayed rectifier-type current. In
order to study the effects of PKA, we compared cell lines transfected with
mKv1.1 alone with lines cotransfected with both mKv1.1 and a plasmid
encoding a dominant negative mutation in the regulatory subunit of PKA.
These mutant regulatory subunits bind to endogenous catalytic subunits of
PKA but do not respond to cAMP, thereby causing a chronic reduction in the
basal PKA activity in these cells. We found that mKv1.1 current kinetics
are unaltered but current density is 3.4-fold higher in the cell lines
expressing mutant regulatory subunit than in lines expressing only mKv1.1.
RNase protection assays indicate that levels of the specific RNA for mKv1.1
are increased almost twofold in the lines expressing mutant regulatory
subunit over the lines expressing mKv1.1 only. Further, the levels of
mKv1.1 protein, assayed using an mKv1.1 channel- specific antibody, are
increased by almost a factor of 3 between the two types of cell lines.
These results suggest that PKA can regulate mKv1.1 channel expression by
changing steady-state levels of RNA and by other posttranscriptional
mechanisms.
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