Positive and negative hepatic regulation of the human type II phospholipase A2 gene.


:To identify the elements which regulate the liver transcription of the human type II phospholipase A2 gene and its stimulation by interleukin 6, the 5' flanking region from -1614 to +806 and several 3' and 5' deleted fragments have been analyzed in CAT assays. Negative regulatory elements have been located in the regions -1614 to -326 and +20 to +806. The fragment -326 to +20 contains the main elements required for the transcription as well as for the stimulation by interleukin 6. Footprinting assays have been performed on this region and showed four protected elements, A [-35;-6], B [-125;-86], C [-209;-176], and D [-247;-211]. Deletion of element D enhanced the transcription of the reporter gene 10.5-fold compared to the [-326;+20]-CAT construct. Further deletions up to position -87 which removed both the elements B and C or the substitution of element C by a nonspecific sequence lowered the promoter activity to 23% and 70% of the control, respectively. These results indicate that element C binds positive regulatory factors and element D binds a negative regulatory factor. Furthermore, stimulation by interleukin 6 is lost when element C is substituted or deleted. As shown by the footprinting and band shift assays, the transcription factors C/EBP alpha and C/EBP beta can bind to elements C and D but the dissociation constant (Kd) of C/EBP alpha is 10 times lower for element C (0.6 nM) than for element D (5.8 nM). Band shift experiments using rat liver nuclear extracts showed that element C formed four heat stable complexes, some of which could be supershifted by anti C/EBP alpha antibodies. The binding of C/EBP factors to element C was confirmed by competition with previously described oligonucleotide and nucleotide substitution of element C. Band shift experiments using rat liver nuclear extracts showed that element D formed one major DNA-protein complex. This complex could be competed out by oligonucleotides containing a cAMP responsive element (CRE) but not by oligonucleotides containing the binding site of C/EBP. However, anti-CREB antibodies did not supershift this complex. Methylation interference experiments showed the involvement of a G nucleotide upstream to the sequence homologous to CRE in the binding of the hepatic nuclear factors.(ABSTRACT TRUNCATED AT 400 WORDS)






Olivier JL,Fan Q,Salvat C,Ziari M,Kong L,Mangeney M,Bereziat G




Has Abstract


1994-06-14 00:00:00












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    pub_type: 杂志文章


    authors: Mitchell DJ,Nikolic D,Jang MH,van Breemen RB,Hille R,Silverman RB

    更新日期:2001-07-24 00:00:00

  • Dissociation of the α-subunit Calf-2 domain and the β-subunit I-EGF4 domain in integrin activation and signaling.

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    pub_type: 杂志文章


    authors: Wang W,Fu G,Luo BH

    更新日期:2010-11-30 00:00:00

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    pub_type: 杂志文章


    authors: Bourdon P,Fenton JW 2nd,Maraganore JM

    更新日期:1990-07-10 00:00:00

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    pub_type: 杂志文章


    authors: Mao B,Gu Z,Gorin A,Chen J,Hingerty BE,Amin S,Broyde S,Geacintov NE,Patel DJ

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    pub_type: 杂志文章,评审


    authors: Kahn JD

    更新日期:2000-04-04 00:00:00

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    pub_type: 杂志文章,评审


    authors: Hammes GG

    更新日期:2002-07-02 00:00:00

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    pub_type: 杂志文章


    authors: Hartman FC

    更新日期:1981-02-17 00:00:00

  • Stimulation of iron(II) bleomycin activity by phosphate-containing compounds.

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    pub_type: 杂志文章


    authors: Burger RM,Horwitz SB,Peisach J

    更新日期:1985-07-02 00:00:00

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    pub_type: 杂志文章


    authors: Carberry SE,Friedland DE,Rhoads RE,Goss DJ

    更新日期:1992-02-11 00:00:00

  • Active site mapping of the serine proteases human leukocyte elastase, cathepsin G, porcine pancreatic elastase, rat mast cell proteases I and II. Bovine chymotrypsin A alpha, and Staphylococcus aureus protease V-8 using tripeptide thiobenzyl ester substra

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    pub_type: 杂志文章


    authors: Harper JW,Cook RR,Roberts CJ,McLaughlin BJ,Powers JC

    更新日期:1984-06-19 00:00:00

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    pub_type: 杂志文章


    authors: Stierl M,Penzkofer A,Kennis JT,Hegemann P,Mathes T

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    authors: Sandberg A,Harrison DJ,Karlsson BG

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    authors: Povirk LF,Hogan M,Dattagupta N

    更新日期:1979-01-09 00:00:00

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    更新日期:1978-05-30 00:00:00

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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    pub_type: 杂志文章


    authors: Pucadyil TJ,Kalipatnapu S,Harikumar KG,Rangaraj N,Karnik SS,Chattopadhyay A

    更新日期:2004-12-21 00:00:00