Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells.
We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution.
Tillman
J B
JB
Department of Biochemistry, University of California, Riverside 92521.
Crone
D E
DE
Kim
H S
HS
Sprung
C N
CN
Spindler
S R
SR
D000818:Animals; D016477:Artifacts; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D001517:Coleoptera / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005822:Genetic Vectors; D007668:Kidney; D008156:Luciferases / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D011988:Receptors, Thyroid Hormone / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D014284:Triiodothyronine / Q000494:pharmacology*
D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D014284:Triiodothyronine; D008156:Luciferases; D013937:Thymidine Kinase