Cloning, functional expression, and regulation of two K+ channels in human T lymphocytes

J Biol Chem. 1992 Apr 25;267(12):8650-7.

Abstract

Low stringency screening of a Jurkat cDNA library with a rat brain K+ channel (RCK1) probe has resulted in the isolation of HLK3, a voltage-gated K+ channel. In Xenopus oocytes, the HLK3 clone directs the expression of a rapidly activating transient outward K+ current similar to the type n K+ current recorded in Jurkat T cells. The HLK3 gene is located on the short arm of human chromosome 1 (p13.3). Polymerase chain reaction was used to clone HIsK from Jurkat cDNA. The HIsK clone shares the same sequence with a previously described genomic clone (Murai, T., Kazikuka, A., Takumi, T., Ohkubo, H., and Nakanishi, S. (1989) Biochem. Biophys. Res. Commun. 161, 176-181). In Xenopus oocytes, it encodes a slowly activating, noninactivating K+ channel which cannot be recorded in Jurkat cells by conventional patch-clamp techniques. Transcripts of both clones are present at a similar level before and after activation of purified human T lymphocytes and Jurkat cells, reflecting a constitutive expression of K+ channel messages. This finding is in good agreement with the electrophysiological results for type n K+ current density on the same cells. HLK3 current is very sensitive to the scorpion toxin charybdotoxin (IC50 = 0.8 nM). HIsK current is totally insensitive to this toxin but is blocked by the antiarrhythmic clofilium (IC50 = 80 microM). While charybdotoxin has no effect on interleukin 2 mRNA induction, clofilium potently inhibits interleukin 2 mRNA expression upon mitogen-induced T cell activation. It is concluded that the HLK3 channel is not an important component of the T cell mitogenic response. Other targets for K+ channel blockers, such as the HIsK protein, could be involved in the activation process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Cell Division / drug effects
  • Charybdotoxin
  • Chromosome Mapping
  • Chromosomes, Human, Pair 1
  • Cloning, Molecular
  • DNA / genetics
  • Gene Expression
  • Humans
  • Interleukin-2 / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Potassium Channels / drug effects
  • Potassium Channels / genetics*
  • Protein Biosynthesis
  • Quaternary Ammonium Compounds / pharmacology
  • RNA, Messenger / genetics
  • Scorpion Venoms / pharmacology
  • Sequence Alignment
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism*
  • Transcription, Genetic
  • Xenopus

Substances

  • Interleukin-2
  • Potassium Channels
  • Quaternary Ammonium Compounds
  • RNA, Messenger
  • Scorpion Venoms
  • Charybdotoxin
  • clofilium
  • DNA

Associated data

  • GENBANK/M74316
  • GENBANK/M74317
  • GENBANK/M74318
  • GENBANK/M74319
  • GENBANK/M74320
  • GENBANK/M74321
  • GENBANK/M74322
  • GENBANK/M74323
  • GENBANK/M74324
  • GENBANK/M85217