A novel aberrant splicing of Gsα transcript in human leukemia cell lines

Abstract

The a subunit of the stimulatory G protein, Gsα, is involved in the stimulation of adenylate cyclase pathway of signal transduction. The various Gsα, mRNAs are generated either by alternative splicing or by using alternative promoter. A novel aberrant splicing of Gsα transcript in human leukemia cell lines is reported. The entire coding region of Gsα gene obtained, referred to as GsαLeu, was amplified by PCR with cDNA from Jurkat human leukemia cDNA library or the reverse-transcribed first strand cDNA from human leukemia cell line HL-60 as a template. The result of DNA sequencing indicated that, compared with originally reported Gsα, GsαLeu has two in-frame deletions, one in amino acid residues 23–249 and the other in amino acid residues 254–260, suggesting that GsαLeu, may play an important role in leukemia. The complete coding region of GsαLeu was inserted downstream of the thrombin site of pGEX-2T fusion protein expression vector and expressed in the form of GST/GsαLeu, at high level, which was then purified to be electrophoretically pure by one-step affinity chromatography. The expression of GsαLeu lays foundation for the study of its function.