Structure and differential mechanisms of regulation of expression of a serine esterase gene in activated human T lymphocytes.
A cDNA clone encoding a polypeptide resembling proteolytic serine esterases (from cytotoxic T-cells; SECT) was isolated from human peripheral blood lymphocytes which had been cultured in the presence of the T-cell mitogen phytohemagglutinin for 72 h. The cDNA encodes a polypeptide of 247 amino acids which show homology of 99% with the protein sequence encoded by a cDNA clone (1-3E) isolated from staphylococcal enterotoxin A-stimulated human peripheral blood lymphocytes and 68% with the protein sequence (cytotoxic cell protease type I) derived from a cDNA clone (C11) encoding a serine esterase isolated from a murine cytotoxic T-cell line. The overall nucleotide sequence homology between the SECT cDNA and 1-3E was 99% and 73% between SECT and C11. Comparing the coding regions of SECT and C11 showed 75% homology, whereas the 5'- and 3'-untranslated regions showed 67% homology. Phytohemagglutinin stimulation results in 30-, 60-, and 370-fold increases in cytoplasmic SECT mRNA with respect to unstimulated cells after 6-, 24-, and 72-h cultures, respectively. At 6 h, the increase in SECT mRNA occurs in the absence of increases in SECT gene transcription and cytoplasmic mRNA stabilization. A 5-fold increase in SECT nuclear RNA seen at this time suggests that stabilization of SECT nuclear RNA transcript is responsible for early increases in SECT mRNA levels. At 24 and 72 h, the increased cytoplasmic SECT mRNA levels can be accounted for by increased transcriptional activity of the SECT gene.
Caputo
A
A
Children's Medical Research Foundation, University of Sydney, Australia.
Fahey
D
D
Lloyd
C
C
Vozab
R
R
McCairns
E
E
Rowe
P B
PB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002478:Cells, Cultured; D002919:Chymotrypsinogen / Q000032:analysis; D004247:DNA / Q000032:analysis; D004950:Esterases / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D013601:T-Lymphocytes / Q000201:enzymology*
D004247:DNA; D002919:Chymotrypsinogen; D004950:Esterases; C022635:serine esterase