Identification of novel mediators of NF-kappaB through genome-wide survey of monocyte adherence-induced genes.
The transcription factor nuclear factor (NF)-kappaB controls the expression of genes involved in inflammation, cell proliferation, apoptosis, and differentiation. Impaired regulation of NF-kappaB has been associated with many diseases; thus, there is significant interest in therapeutic approaches based on modulation of this transcription factor. NF-kappaB activity is controlled by numerous signaling molecules, many of which are potentially to be identified. Monocytes are principal effectors of the immune system, and monocyte adherence is the first step leading to their activation and differentiation. Adherence induces activation of NF-kappaB, resulting in the induction of proinflammatory genes as well as anti-inflammatory genes, which counterbalance and limit the intensity and duration of NF-kappaB activation. Here, to identify novel mediators of NF-kappaB signaling, we used the model of monocyte adherence to perform a systematic, genome-wide survey of adherence-induced genes. Having isolated mRNAs from nonadherent and adherent primary human monocytes, we constructed suppressive subtraction hybridization libraries containing cDNAs, which were differentially regulated by adherence. Of 366 identified differentially expressed genes, most were found to be up-regulated by adherence. Having analyzed a subset of these genes, we found that the library was enriched with inhibitors of NF-kappaB. Three of those (an orphan nuclear receptor NUR77, a guanosine 5'-diphosphate/guanosine 5'-triphosphate exchange factor RABEX5, and a PRK1-associated protein AWP1) were particularly potent inhibitors of NF-kappaB activation. Thus, the collection of monocyte adherence-regulated genes represents a rich source for the identification of novel components of the machinery that controls NF-kappaB activation.
Diatchenko
Luda
L
Attagene Inc., 7030 Kit Creek Road, Research Triangle Park, NC 27560, USA.
Romanov
Sergei
S
Malinina
Inga
I
Clarke
Julie
J
Tchivilev
Igor
I
Li
Xiangli
X
Makarov
Sergei S
SS
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D002448:Cell Adhesion / Q000235:genetics; D002478:Cells, Cultured; D002634:Chemotaxis, Leukocyte / Q000235:genetics* / Q000276:immunology; D004268:DNA-Binding Proteins / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics*; D005796:Genes / Q000235:genetics; D016678:Genome / Q000235:genetics*; D015698:Genomic Library; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000276:immunology; D009000:Monocytes / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D011987:Receptors, Steroid / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D014157:Transcription Factors / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; D020662:Guanine Nucleotide Exchange Factors; D016328:NF-kappa B; C539821:NR4A1 protein, human; D057105:Nuclear Receptor Subfamily 4, Group A, Member 1; C108382:RABGEF1 protein, human; D018160:Receptors, Cytoplasmic and Nuclear; D011987:Receptors, Steroid; D014157:Transcription Factors; C417945:ZFAND6 protein, human