Molecular and in silico characterization of a promoter module and C/EBP element that mediate LPS-induced RANTES/CCL5 expression in monocytic cells

FASEB J. 2001 Mar;15(3):577-9. doi: 10.1096/fj.00-0459fje. Epub 2001 Jan 5.


The chemokine RANTES/CCL5 is a proinflammatory agent produced by a variety of tissues in response to specific stimuli. In human monocytes, RANTES/CCL5 transcription is up-regulated rapidly and transiently in response to LPS. We describe here two regions that help control LPS-driven transcription from the human RANTES/CCL5 promoter in monocytic cells. These sites were analyzed by using DNase I footprinting, transient transfection assays, site-directed mutagenesis, and EMSA. RANTES site E (R(E), -125/-99) constitutively binds C/EBP proteins in monocytic Mono Mac 6 cells. Mutation of region R(E) led to a significant (40%-50%) reduction in LPS-induced promoter reporter activity. Region R(AB) is composed of tandem kB-like elements R(A) and R(B) (-73/-34). These sites working in concert act as an LPS-responsive promoter module. R(A) constitutively binds Sp1, and Rel p50/p65 following LPS stimulation. Either factor can mediate transcriptional effects at R(A). Induced Rel p50/p50 binding to site R(B) is required for LPS regulation of RANTES/CCL5 transcription. A series of computer models based on the RANTES/CCL5 promoter were generated to represent the organization of these functional elements. The models could identify LPS-regulated promoters in human, other vertebrate, and viral sequences in various databases.

MeSH terms

  • CCAAT-Enhancer-Binding Proteins / metabolism*
  • Cell Line
  • Chemokine CCL5 / genetics
  • Chemokine CCL5 / metabolism*
  • Computer Simulation
  • Dimerization
  • Genes, Reporter / genetics
  • Humans
  • Lipopolysaccharides / pharmacology*
  • Monocytes / drug effects
  • Monocytes / metabolism*
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Promoter Regions, Genetic / genetics*
  • Sp1 Transcription Factor / genetics
  • Sp1 Transcription Factor / metabolism
  • Transfection


  • CCAAT-Enhancer-Binding Proteins
  • Chemokine CCL5
  • Lipopolysaccharides
  • NF-kappa B
  • Sp1 Transcription Factor