Ag. Sarafanov et al., Human tissue plasminogen activator gene: Regulation of expression by signals of the 3 '-untranslated region, MOL BIOL, 33(3), 1999, pp. 360-369
Constructs consisting of a reporter chloramphenicol acetyltransferase (CAT)
gene and various deletion versions of the 3'-untranslated region (3UTR) of
the human tissue plasminogen activator (tPA) gene were transiently express
ed in mammalian cells. In all cases, 3UTR of the tPA gene inhibited the CAT
gene expression five- to tenfold, suggesting similar mechanisms of the eff
ect in different mammalian tissues. Experiments on the expression of the ne
sted 3UTR deletion constructs in the CV1 cells allowed one to localize thre
e regions (83, 169, and 219 bp) with signals influencing the reporter gene
expression. The first region increased the expression 1.5-2 times, while th
e second and the third ones inhibited it 2.5-3 times each. Computer analysi
s revealed substantial homology of these regions with 3UTRs of the mouse an
d rat tPA genes and their localization in the distinct hairpin-like regions
of the mRNA predicted secondary structure. On the basis of pCAT-Control, a
n expression vector permitting examination of the 3UTR regulatory elements
of different genes was constructed.