Two regions within the human IL-2 gene promoter are important for inducible IL-2 expression.
We have examined regulatory domains of the human IL-2 gene promoter by transfection and transient expression of rDNA constructs in which the chloramphenicol acetyl transferase gene shows T cell-specific inducible expression and cyclosporin A-mediated inhibition when placed downstream of 587 bp of the human IL-2 5'-flanking region. A series of 5'-deletion constructs transfected into the Jurkat T lymphoid line demonstrates that a region encompassing 370 bp 5' of the transcription start site is sufficient for inducible chloramphenicol acetyl-transferase expression. Further dissection of this region with internal deletion (linker-scanner) mutants revealed that portions of at least two discrete regions from -42 to -169 and -289 to -361 bp relative to the transcription start site are critical for inducible expression of the IL-2 gene. T cell-specific expression of wild-type and mutant IL-2 promoter constructs could be increased severalfold by the insertion of an upstream SV40 enhancer. With use of a battery of IL-2 promoter constructs, we could not identify subregions within IL-2 5'-flanking sequences which are crucial for cyclosporin A inhibition of the IL-2 gene or deletion of which resulted in loss of T cell-specific expression, suggesting that such functions may be mediated at pre-transcriptional levels.
Williams
T M
TM
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
Eisenberg
L
L
Burlein
J E
JE
Norris
C A
CA
Pancer
S
S
Yao
D
D
Burger
S
S
Kamoun
M
M
Kant
J A
JA
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D001402:B-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase; D002872:Chromosome Deletion; D003524:Cyclosporins / Q000494:pharmacology; D005796:Genes / Q000187:drug effects; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000187:drug effects; D013539:Simian virus 40 / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D014162:Transfection
D003524:Cyclosporins; D007376:Interleukin-2; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase