Involvement of two NF-kappaB binding sites in PMA-induced expression of the human leukotactin-1/CCL15 gene in U937 monocytoid cells.
Leukotactin-1 (Lkn-1)/CCL15, is a recently cloned chemotactic chemokine that appears to play important roles in the inflammatory process by recruiting immune cells to inflammatory sites. Expression of the Lkn-1/CCL15 gene is inducible in monocytes but its transcriptional regulation has not been studied. To identify Lkn-1/CCL15 regulatory sequences in monocytic cells, U937 cells were transiently transfected with the luciferase reporter gene linked to various deletions of the Lkn-1/CCL15 promoter region. The region -269 to -43 bp from the transcription start site proved to be important for induction by PMA. This region contained two potential NF-kappaB sites: one between -191 and -182 bp, and the other between -60 and -51 bp. Mutation of either element reduced PMA-induced expression and electrophoretic mobility shift assays revealed that NF-kappaB recognized both potential NF-kappaB sites. In addition, PMA-induction of Lkn-1/CCL15 in transiently transfected U937 cells was blocked by proteasome inhibitor 1. These observations demonstrate that the two NF-kappaB binding sites are essential for PMA-induced Lkn-1/CCL15 expression in human monocytes.
Shin
Yong-Hyun
YH
Graduate School of Biotechnology, Institute of Life Sciences and Resources, Kyung Hee University, Yongin 449-701, Korea.
Shim
Jung-Jae
JJ
Hur
Man-Wook
MW
Kang
Chang-Joong
CJ
Kim
Jiyoung
J
D001665:Binding Sites; D002460:Cell Line; D019742:Chemokines, CC / Q000235:genetics* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D009000:Monocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D016328:NF-kappa B / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D019742:Chemokines, CC; D004791:Enzyme Inhibitors; D016328:NF-kappa B; D013755:Tetradecanoylphorbol Acetate