Transcriptional activation of alpha 1(III) procollagen gene in Tsk2/+ dermal fibroblasts.
Transient transfection experiments into Tsk2/+ and normal dermal fibroblasts were performed using four successively shorter Col3a1 promoter deletion constructs: #103, #110, #114, and #120 fused to the chloramphenicol-acetyl-transferase (CAT) reporter gene. The transcriptional activity in Tsk2/+ and normal dermal fibroblasts driven by the three longer constructs was equal. With the shortest construct, #120 (-96 to +16bp) the transcriptional activity in Tsk2/+ fibroblasts was 25 times higher than in normal fibroblasts. Electrophoretic mobility shift assays with a labeled #120 probe revealed that increased DNA-protein binding occurred with nuclear extracts prepared from Tsk2/+ fibroblasts and that this binding was displaced by consensus Sp1 and NF-1 oligonucleotide sequences. These data indicate that sequences from -96 to +16bp of the Col3a1 promoter play an important role in the upregulated expression of this gene in Tsk2/+ fibroblasts and that the promoter contains sequences which bind the trans-acting nuclear factors, Sp1(like) and NF-1(like).
Christner
Paul J
PJ
Division of Rheumatology, Department of Medicine, Jefferson Medical College, Thomas Jefferson University, Room 509, Bluemle Life Sciences Building, 233 South 10th Street, Philadelphia, PA 19107-5541, USA. Paul.Christner@mail.tju.edu
Yufit
Tatyana
T
Peters
Josephine
J
McGrath
Rodney
R
Conway
Raymond F
RF
Jiménez
Sergio A
SA
D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D024061:Collagen Type III / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection
C439971:COL3A1 protein, human; D024061:Collagen Type III; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase