Detection of a functional promoter/enhancer in an intron-less human gene encoding a glutamine synthetase-like enzyme.
A human genomic clone, psi GS, containing an intron-less glutamine synthetase (GS)-encoding pseudogene, was isolated by screening a human library. A sequence of 3004 bp, containing the GS coding region and both the 5' and 3' flanking sequences, was identified that exhibits all the characteristics of a processed pseudogene. The coding region shows 93% identity with the human GS cDNA (hGS) sequence and contains two frame-shifts and two termination codons. The coding sequence is flanked by a 9-bp AT repeat and a putative polyadenylation site, AATAAA, at the 3' end. Primer extension analysis and S1 nuclease mapping showed a transcription start point (tsp) 62 bp upstream from the start codon indicating a shorter untranslated region than hGS. Transfection of HeLa cells with cat constructs containing portions of the 5' flanking sequence showed the presence of a functional promoter/enhancer within 200 bp of the tsp, independent of its orientation.
Chakrabarti
R
R
Department of Surgery, University of Florida, Gainesville 32610.
McCracken
J B
JB
Chakrabarti
D
D
Souba
W W
WW
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005974:Glutamate-Ammonia Ligase / Q000737:chemistry / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000166:cytology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011544:Pseudogenes / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic / Q000235:genetics
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D005974:Glutamate-Ammonia Ligase