Negative-Dominant IB Kinase inhibits NF-B and APRE Reporter Plasmids
A. Arman
The New England Baptist Bone & Joint Institute, Harvard Institute of Medicine, Harvard Medical
School, Harvard University, 77 Avenue Louis Pasteur, Boston, MA 02115l; Marmara University, The
Faculty of Engineering, Environmental Engineering, Biotechnology Group, Molecular Biology and
Genetics and Cell Culture Lab. Room # 405, Goztepe Campus, 34 744 Istanbul, Turkey,
Phone: 90 216 348-0292/608; Fax: 90 216 3450126; e-mail: aarman@eng.marmara.edu.tr
Interleukin 1 (IL-1) is a multifunctional pro-inflammatory factor that is involved in fever, increased
acute phase response, metastases, angiogenesis, cartilage breakdown, cell differentiation. It is existing
two active forms, IL-1 and IL-1. Also, IL-1R antagonist (IL-1Ra) form exists in circulation. IL-1
or  acts as agonist but IL-1Ra acts as antagonist. IL-1 plays an important role mediating
inflammatory diseases such as rheumatoid arthritis, coroner arter diseases. IL-1 shows these activities
through IL-1 receptor (IL-1R) and another IL-1R-like molecule (IL-1Racp). This ternary complex is
required to start the initiation of signaling. IL-1 initiates at least 3 signal transduction pathways which
are IL-1-induced IRAK pathway, IL-1-induced PI3-kinase pathway and IL-1-induced JAK-STAT
pathway. Recent studies demonstrated that IB kinases regulates NF-B proteins. The NF-B
proteins are family of transcription factors that consists of 5 members which are p65 (RelA), RelB, cRel, NF-B1 and NF-B2. NF-B transcription factors are localized in cytoplasm and are activated by
ligands such as IL-1. After the activation of NF-B proteins, they enter nucleus and bind to promoter
of the gene and activate gene expression.
Aim: Purpose of this research is to determine the effect of IB kinase on NF-B and APRE Reporter
Plasmids.
Methods: Transfection. Hep3 cells were seeded in 12 well-plates and cotransfeceted dominantnegative IB Kinase plasmid with APRE or NF-B reporter plasmids. After 48 hours later, the cells
were treated with or without IL-1 (1ng/ml) for 5 hours and then the cells were harvested and luciferase
activities of cells were measured.
Results and Discussion: Transient-transfection assay results show that negative dominant IB Kinase
caused the reduction on the APRE and NF-B reporter activities. This implies that Stat3 and NF-B
transcription factors may interact each other at APRE promoter and activate the gene activation.