Identification of a Zinc Finger Protein that Inhibits IL-2 Gene Expression

Williams, Tiffani L.; Moolten, David; Burlein, Jeanne E.; Romano, Josephine; Bhaerman, R; Godillot, Alexis P.; Mellon, Mark T.; Rauscher, F. J.; Kant, JA

doi:10.1126/science.1840704

Cited by 180 publications

(100 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results demonstrated that the optimal consensus binding sequence of AREB6#2 is C/TACCTG/TT. This matches the inverted sequence of Nil-2-a binding site, NRE-A(GA-CAGGTAAA), which was identified as the negative regulatory element located upstream of the IL-2 gene promoter (Williams et al, 1991). To determine the optimal binding sequences or detailed binding sequence requirements for AREB6#1 and AREB6#5 in comparison with AREB6#2, we performed gel mobility shift assays with the same set of competitors as shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…As the second set, we performed transient cotransfection into Jurkat cells using the naturally occurring IL-2 promoter, which contains the Nil-2-a binding site, NRE, at -110 to -101 (Williams et al, 1991). We observed that only AREB6CZ, not AREBGNZ, binds to the element in gel mobility shift assays (data not shown).…”

Section: F 1 1 : T a T C A T G A C C C C A C C C A C G A T A A G A T mentioning

confidence: 99%

“…that the AREB6 cDNA clone corresponds to a 5'-extended version of Nil-2-a originally isolated as a negative regulator of the interleukin 2 (IL-2) gene (Williams et al, 1991). Furthermore, AREB6 is a human homolog of chicken 6EF1, which binds to the enhancer element of chicken 61-crystallin gene and represses its expression (Funahashi et al, 1993).…”

Section: Sequence Analysis Indicatesmentioning

confidence: 99%

See 2 more Smart Citations

DNA Binding through Distinct Domains of Zinc‐Finger‐Homeodomain Protein AREB6 has Different Effects on Gene Transcription

Ikeda

Kawakami

1995

European Journal of Biochemistry

View full text Add to dashboard Cite

Transcription factor AREB6 has a unique structure composed of two zinc-finger clusters in N-and C-terminal regions, and one homeodomain in the middle. AREB6 has been known to regulate the expression of the Na, K-ATPase a1 subunit, interleukin 2 and &crystallin genes. We determined the optimal binding sites for the N-terminal zinc-finger cluster as GTCACCTGT or TGCACCTGT and for the Cterminal zinc-finger cluster as C/TACCTG/TT by the CASTing method (cyclic amplification and selection of targets). The additional consensus sequence GTTTC/G, in conjunction with the CACCTGT sequence, was selected by the second CASTing for the entire coding region. The N-terminal zinc-finger cluster binds to DNA strongly when the DNA has GTTTC/G in conjunction with the CACCTGT sequence. The homeodomain had no specific DNA binding activity but was found to interact with the N-terminal zincfinger cluster. Analyses of zinc-finger mutation proteins revealed that the contribution to DNA binding of each N-terminal zinc-finger motif is altered depending on the presence of the additional consensus. Transient transfection assays showed that AREB6 repressed the human 70-kDa heat-shock gene promoter harboring the CACCTGT sequence together with the additional consensus, and that AREB6 activated the promoter harboring the CACCTGT sequence without the additional consensus. These results suggest that AREB6 has multiple conformational states, leading to positive and negative regulations of gene transcription.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: F 1 1 : T a T C A T G A C C C C A C C C A C G A T A A G A T mentioning

confidence: 99%

Section: Sequence Analysis Indicatesmentioning

confidence: 99%

See 1 more Smart Citation

DNA Binding through Distinct Domains of Zinc‐Finger‐Homeodomain Protein AREB6 has Different Effects on Gene Transcription

Ikeda

Kawakami

1995

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…A negative regulatory element seems important for repression of IL-2 gene transcription (61), and expression of this inhibitory factor in unstimulated Jurkat T cells may explain the lack of NFATc activity on the intact IL-2 gene promoter (Fig. 5).…”

Section: Egr-1 Regulates Il-2 Transcriptionmentioning

confidence: 99%

The Early Growth Response Protein (EGR-1) Regulates Interleukin-2 Transcription by Synergistic Interaction with the Nuclear Factor of Activated T Cells

Decker

Skerka

Zipfel

1998

Journal of Biological Chemistry

View full text Add to dashboard Cite

The early growth response-1 gene (EGR-1) is induced by a wide range of stimuli in diverse cell types; however, EGR-1-regulated genes display a highly restricted pattern of expression. Recently, an overlapping Sp1⅐EGR-1 binding site has been identified within the interleukin-2 (IL-2) gene promoter directly upstream of the binding site for the nuclear factor of activated T cells (NFAT). We used transfection assays to study how the abundantly and constitutively expressed Sp1 protein and the immediate early EGR-1 zinc finger protein regulate IL-2 gene expression. Here, we identify EGR-1 as an important activator of the IL-2 gene. In Jurkat T cells, EGR-1 but not Sp1 acts as a potent coactivator for IL-2 transcription, and in combination with NFATc, EGR-1 increases transcription of an IL-2 reporter construct 200-fold. Electrophoretic mobility shift assays reveal that recombinant EGR-1 and NFATc bind independently to their target sites within the IL-2 promoter, and the presence of both sites on the same DNA molecule is required for EGR-1⅐NFATc⅐DNA complex formation. The transcriptional synergy observed here for EGR-1 and NFATc explains how the abundant nuclear factor EGR-1 contributes to the expression of restrictively expressed genes.

show abstract

“…For the assay of AP-1 activity, nuclear protein extract (10 pg) was incubated with 1.5 pg poly(d1-dC) in 12 mM Hepes pH 7.9, 60 mM KCI and 12% glycerol for 15 rnin at room temperature (Williams et al, 1991). After addition of "P-labelled AP-1 binding oligonucleotide (10" cpm), the mixture was incubated for 15 min at room temperature and was then loaded on a native 6% polyacrylamide gel (acrylamide/bisacrylamide 49 : 1) containing 40 mM Tris, 226 mM glycine pH 8.2 (Caceres et al, 1991).…”

Section: Electrophoretic Mobility Shift Assaymentioning

confidence: 99%

Distinct effects of glutathione disulphide on the nuclear transcription factors κB and the activator protein‐1

Galter

Mihm

Dröge

1994

European Journal of Biochemistry

263

144

View full text Add to dashboard Cite

Oxidative conditions potentiate the activation of the nuclear transcription factor KB (NFKB) and the activator protein-1 (AP-1) in intact cells, but inhibit their DNA binding activity in vitro. We now show that both the activation of NFKB and the inhibition of its DNA binding activity is modulated in intact cells by the physiological oxidant glutathione disulphide (GSSG). NFKB activation in human T lineage cells (Molt-4) by 12-0-tetradecanoyl-phorbol 13-acetate was inhibited by dithiothreitol, and this was partly reversed by the glutathione reductase inhibitor 1,3-bis(2-chloroethyl)-1 -nitrosourea (BCNU) or by hydrogen peroxide, indicating that GSSG may be required for NFKB activation. These effects of BCNU and hydrogen peroxide were not seen in glutathionedepleted cells. However, NFKB and AP-1 activation were potentiated by dithiothreitol if added to cell cultures 1 h after the phorbol ester, indicating that a shift of redox conditions may support optimal oxidative activation with minimal inhibition of DNA binding. The elevation of intracellular GSSG levels by BCNU before stimulation suppressed the chloramphenicol acetyltransferase expression dependent on NFKB but increased that dependent on AP-1. This selective suppression of NFKB was also demonstrable by electrophoretic mobility shift assays. In vitro, GSSG inhibited the DNA binding activity of NFKB more effectively than that of AP-1, while AP-3 was inhibited more effectively by oxidized thioredoxin.The nuclear transcription factor KB (NFKB) is involved in the inducible transcription of immunologically relevant genes including the genes of the interleukin-2 receptor achain, tumor necrosis factor-a, major histocompatibility complex antigens and c-fos (Leung and Nabel, 1988;Cross et al., 1989; reviewed by Ullmann et al., 1990). The NFKB protein is found in many different cell types including B and T lymphocytes, macrophages and monocytes (reviewed by Baeuerle and Baltimore, 1991). The transcriptional activation of NFKB-dependent genes is usually induced by dissociation of the inhibitory protein IKB from the NFKB heterodimer in the cytoplasm and the subsequent translocation of active NFKB into the nucleus. The Fos and Jun proteins are components of another family of nuclear transcription factors that have been implicated in the regulation of growth, differentiation, neuronal excitation and cellular stress in a number of different cell types (reviewed by Angel and Karin, 1991;. Fos and Jun form a heterodimeric complex that interacts with a DNA sequence known as the activator protein-1 (AP-1) binding site or TPA-responsive element (TRE; TPA = 12-0-tetradecanoyl-phorbol 13-acetate). NFKB and other related factors of the v-and c-Re1 oncoprotein family share a characteristic sequence motif with a cysteine and three arginine residues in the DNA binding region (Ghosh et al., 1990;Kieran et al., 1990;Kumar et al., 1992). The DNA binding region of Fos, Jun and several related transcription factors also contains a characteristic sequence with a cysteine and up to seven ar...

show abstract

Identification of a Zinc Finger Protein that Inhibits IL-2 Gene Expression

Cited by 180 publications

References 33 publications

DNA Binding through Distinct Domains of Zinc‐Finger‐Homeodomain Protein AREB6 has Different Effects on Gene Transcription

DNA Binding through Distinct Domains of Zinc‐Finger‐Homeodomain Protein AREB6 has Different Effects on Gene Transcription

The Early Growth Response Protein (EGR-1) Regulates Interleukin-2 Transcription by Synergistic Interaction with the Nuclear Factor of Activated T Cells

Distinct effects of glutathione disulphide on the nuclear transcription factors κB and the activator protein‐1

Contact Info

Product

Resources

About