A novel transiently expressed, integral membrane protein linked to cell activation. Molecular cloning via the rapid degradation signal AUUUA.
A novel cDNA clone termed E16 which codes for an integral membrane protein of 241 amino acids with six transmembrane domains was isolated from peripheral blood lymphocytes. The cDNA clone is 4000 base pairs in length and exhibits an unusually long 3'-untranslated region of about 3000 nucleotides. Its expression at the mRNA level is closely linked to cellular activation and division. In all myeloid and lymphoid cells, as well as in primary lymphocytes from peripheral blood, E16 transcripts are rapidly induced and rapidly degraded after stimulation. This pattern of expression is unusual for an integral membrane protein and resembles more closely the kinetic seen for protooncogenes and lymphokines in the T cell system. Its isolation was made possible by a novel approach especially designed to selectively clone cDNAs which exhibit such an expression kinetic. It is based on a combination of the differential screening of a subtracted cDNA library and the subsequent hybridization of the resulting phages to a short oligonucleotide (5'-TAAATAAA-TAAATA-3'). This oligonucleotide is complementary to a trimer of the rapid degradation signal (AUUUA) which is present as a single or reiterated motif in the 3'-untranslated region of many short-lived transcripts.
Gaugitsch
H W
HW
Department of Immunopharmacology, Sandoz Research Institute Ltd., Vienna, Austria.
Prieschl
E E
EE
Kalthoff
F
F
Huber
N E
NE
Baumruker
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D016207:Cytokines / Q000235:genetics; D004247:DNA / Q000235:genetics; D015759:Ionomycin / Q000494:pharmacology; D007700:Kinetics; D008214:Lymphocytes / Q000187:drug effects; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011519:Proto-Oncogenes; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D016207:Cytokines; D008565:Membrane Proteins; D012333:RNA, Messenger; D015759:Ionomycin; D004247:DNA; D013755:Tetradecanoylphorbol Acetate