Upstream NF-kappa B site is required for the maximal expression of mouse inducible nitric oxide synthase gene in interferon-gamma plus lipopolysaccharide-induced RAW 264.7 macrophages

被引:144
作者
Kim, YM [1 ]
Lee, BS [1 ]
Yi, KY [1 ]
Paik, SG [1 ]
机构
[1] CHUNGNAM NATL UNIV,DEPT BIOL,TAEJON 305764,SOUTH KOREA
关键词
D O I
10.1006/bbrc.1997.7031
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Transient transfection assays with various deletion mutants of the mouse inducible nitric oxide synthase (iNOS) promoter linked to a CAT reporter gene demonstrated that, besides the downstream NF-kappa B site, the region from -973 to -925 which contains a potential binding site for NF-kappa B (upstream NF-kappa B site) also mediated lipopolysaccharide (LPS)-inducibility in mouse macrophage cell line RAW 264.7. Site-specific mutation of three conserved nucleotides within the upstream NF-kappa B site abolished additional induction by LPS as well as maximal expression of iNOS by IFN-gamma plus LPS. In contrast, site-specific mutation of the downstream NF-kappa B site caused almost all reduction in expression of the reporter gene by LPS or LPS plus IFN-gamma. Electrophoretic mobility shift assays with the two NF-kappa B sites showed LPS-induced NF-kappa B binding to both probes and its higher affinity to the upstream NF-kappa B site. Taken together, these suggest that the upstream NF-kappa B Site having enhancer function, besides the downstream NF-kappa B site as a core promoter, is essential for maximal expression of the iNOS gene. (C) 1997 Academic Press.
引用
收藏
页码:655 / 660
页数:6
相关论文
共 22 条
[1]  
ALLY EW, 1995, GENE, V158, P247
[2]  
[Anonymous], 1976, ANAL BIOCHEM, V72, P248
[3]   NF-KAPPA-B AND RELATED PROTEINS - REL DORSAL HOMOLOGIES MEET ANKYRIN-LIKE REPEATS [J].
BLANK, V ;
KOURILSKY, P ;
ISRAEL, A .
TRENDS IN BIOCHEMICAL SCIENCES, 1992, 17 (04) :135-140
[4]   ACCURATE TRANSCRIPTION INITIATION BY RNA POLYMERASE-II IN A SOLUBLE EXTRACT FROM ISOLATED MAMMALIAN NUCLEI [J].
DIGNAM, JD ;
LEBOVITZ, RM ;
ROEDER, RG .
NUCLEIC ACIDS RESEARCH, 1983, 11 (05) :1475-1489
[5]   An interferon-gamma-activated site (GAS) is necessary for full expression of the mouse iNOS gene in response to interferon-gamma and lipopolysaccharide [J].
Gao, JJ ;
Morrison, DC ;
Parmely, TJ ;
Russell, SW ;
Murphy, WJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (02) :1226-1230
[6]   TRANSCRIPTIONAL INHIBITION OF THE INDUCIBLE NITRIC-OXIDE SYNTHASE GENE BY COMPETITIVE-BINDING OF NF-KB/REL PROTEINS [J].
GOLDRING, CEP ;
NARAYANAN, R ;
LAGADEC, P ;
JEANNIN, JF .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1995, 209 (01) :73-79
[7]  
Gorman C., 1985, DNA CLONING PRACTICA, V1, P143
[8]   RECOMBINANT GENOMES WHICH EXPRESS CHLORAMPHENICOL ACETYLTRANSFERASE IN MAMMALIAN-CELLS [J].
GORMAN, CM ;
MOFFAT, LF ;
HOWARD, BH .
MOLECULAR AND CELLULAR BIOLOGY, 1982, 2 (09) :1044-1051
[9]  
GRILLI M, 1993, INT REV CYTOL, V143, P1
[10]   THE INDUCIBLE TRANSCRIPTION FACTOR NF-KAPPA-B - STRUCTURE-FUNCTION RELATIONSHIP OF ITS PROTEIN SUBUNITS [J].
GRIMM, S ;
BAEUERLE, PA .
BIOCHEMICAL JOURNAL, 1993, 290 :297-308