Binding of two nuclear complexes to a novel regulatory element within the human S100A9 promoter drives the S100A9 gene expression.
S100A9, also referred to as MRP14, is a calcium-binding protein whose expression is tightly regulated during differentiation of myeloid cells. The present study was performed to study the cell type- and differentiation-specific transcriptional regulation of the S100A9 gene. Analysis of the S100A9 promoter in MonoMac-6 cells revealed evidence for a novel regulatory region from position -400 to -374 bp, termed myeloid-related protein regulatory element (MRE). MRE deletion resulted in a 5.2-fold reduction of promoter activity. By electrophoretic mobility shift analysis two nuclear complexes binding to this region were identified and referred to as MRE-binding complex A (MbcA) and MRE-binding complex B (MbcB). By mutagenesis the MRE-binding motif could be narrowed to a 12-bp region. The relevance of MRE is deduced from the observations that the formation of either MRE-binding complex A or MRE-binding complex B strongly correlated with S100A9 gene expression in a cell type-specific, activation- and differentiation-dependent manner. Moreover, DNA affinity chromatography and Western blot studies indicate that a Kruppel-related zinc finger protein and the transcriptional intermediary factor 1beta (TIF1beta) are involved in an MRE-binding complex, thereby regulating the S100A9 gene expression.
Kerkhoff
Claus
C
Institute of Experimental Dermatology, 48149 Muenster, Germany. kerkhoc@uni-muenster.de
Hofmann
Heiko A
HA
Vormoor
Josef
J
Melkonyan
Harutyun
H
Roth
Johannes
J
Sorg
Clemens
C
Klempt
Martin
M
D001483:Base Sequence; D001853:Bone Marrow / Q000378:metabolism; D040502:Calgranulin B / Q000235:genetics*; D002454:Cell Differentiation; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000502:physiology; D005786:Gene Expression Regulation; D005809:Genes, Regulator / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000502:physiology; D000076183:Tripartite Motif-Containing Protein 28; D016335:Zinc Fingers / Q000502:physiology
D040502:Calgranulin B; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; C101116:TRIM28 protein, human; D000076183:Tripartite Motif-Containing Protein 28