Cloning and expression of the delayed-rectifier IsK channel from neonatal rat heart and diethylstilbestrol-primed rat uterus.
cDNAs encoding a delayed-rectifier-type K+ channel were cloned from both neonatal rat heart and ovariectomized, diethylstilbestrol-primed rat uterus by using the polymerase chain reaction. Both clones have nucleotide sequences identical to that encoding the rat kidney IsK channel [Takumi, T., Ohkubo, H. & Nakanishi, S. (1988) Science 242, 1042-1045] and encode a putative protein of 130 amino acids. Injection of RNA transcripts of the cDNAs into Xenopus oocytes resulted in the expression of a slowly activating, voltage-dependent K+ current. An antisense oligonucleotide, derived from the sequence of the clone, specifically inhibited the expression of the slow, outward current observed in cells injected with mRNAs isolated from the parent tissues (i.e., kidney, heart, and uterus), indicating that the cloned gene underlies the major K+ current expressed from RNA isolated from these tissues.
Folander
K
K
Department of Pharmacology, Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Smith
J S
JS
Antanavage
J
J
Bennett
C
C
Stein
R B
RB
Swanson
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D003001:Cloning, Molecular; D004054:Diethylstilbestrol / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006321:Heart / Q000502:physiology*; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000187:drug effects / Q000502:physiology; D015345:Oligonucleotide Probes; D009865:Oocytes / Q000502:physiology; D010052:Ovariectomy; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D014158:Transcription, Genetic; D014599:Uterus / Q000187:drug effects / Q000502:physiology*; D014982:Xenopus laevis
D008565:Membrane Proteins; D015345:Oligonucleotide Probes; D015221:Potassium Channels; D012333:RNA, Messenger; D004054:Diethylstilbestrol