Binding of the Wilms' tumor locus zinc finger protein to the EGR-1 consensus sequence

Science ◽  
1990 ◽  
Vol 250 (4985) ◽  
pp. 1259-1262 ◽  
Author(s):  
F. Rauscher ◽  
J. Morris ◽  
O. Tournay ◽  
D. Cook ◽  
T Curran
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Wilms Tumor ◽  
Consensus Sequence ◽  
Finger Protein

Two homologous zinc finger genes identified by multicopy suppression in a SNF1 protein kinase mutant of Saccharomyces cerevisiae

1993 ◽  
Vol 13 (7) ◽  
pp. 3872-3881
Author(s):  
F Estruch ◽  
M Carlson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Wilms Tumor ◽  
Temperature Sensitive ◽  
Carbon Utilization ◽  
Functional Roles ◽  
Multicopy Suppressor ◽  
Finger Protein

The MSN2 gene was selected as a multicopy suppressor in a temperature-sensitive SNF1 protein kinase mutant of Saccharomyces cerevisiae. MSN2 encodes a Cys2His2 zinc finger protein related to the yeast MIG1 repressor and to mammalian early growth response and Wilms' tumor zinc finger proteins. Deletion of MSN2 caused no phenotype. A second similar zinc finger gene, MSN4, was isolated, and deletion of both genes caused phenotypic defects related to carbon utilization. Overexpression of the zinc finger regions was deleterious to growth. LexA-MSN2 and LexA-MSN4 fusion proteins functioned as strong transcriptional activators when bound to DNA. Functional roles of this zinc finger protein family are discussed.

scholarly journals A Myc-associated zinc finger protein binding site is one of four important functional regions in the CD4 promoter.

1995 ◽  
Vol 15 (6) ◽  
pp. 3179-3186 ◽  
Author(s):  
D D Duncan ◽  
A Stupakoff ◽  
S M Hedrick ◽  
K B Marcu ◽  
G Siu
Keyword(s):  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Consensus Sequence ◽  
Developmental Expression ◽  
Critical Element ◽  
Factor Binding ◽  
Functional Regions ◽  
Finger Protein ◽  
Myc Gene ◽  
Predominant Factor

The CD4 promoter plays an important role in the developmental control of CD4 transcription. In this report, we show that the minimal CD4 promoter has four factor binding sites, each of which is required for full function. Using biochemical and mutagenesis analyses, we determined that multiple nuclear factors bind to these independent sites. We determined that an initiator-like sequence present at the cap site and an Ets consensus sequence are required for full promoter function. We also demonstrate that the Myc-associated zinc finger protein (MAZ) appears to be the predominant factor binding to one of these sites. This last site closely resembles the ME1a1 G3AG4AG3 motif previously shown to be a critical element in the P2 promoter of the c-myc gene. We therefore believe that the MAZ transcription factor is also likely to play an important role in the control of developmental expression of the CD4 gene.

Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1

1994 ◽  
Vol 14 (3) ◽  
pp. 1979-1985
Author(s):  
M Lundin ◽  
J O Nehlin ◽  
H Ronne
Keyword(s):  
Dna Binding ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Wilms Tumor ◽  
Glucose Repression ◽  
Saturation Mutagenesis ◽  
Sequence Specificity ◽  
Rich Region ◽  
Finger Protein ◽  
Gc Box

MIG1 is a zinc finger protein that mediates glucose repression in the yeast Saccharomyces cerevisiae. MIG1 is related to the mammalian Krox/Egr, Wilms' tumor, and Sp1 finger proteins. It has two fingers and binds to a GCGGGG motif that resembles the GC boxes recognized by these mammalian proteins. We have performed a complete saturation mutagenesis of a natural MIG1 site in order to elucidate its binding specificity. We found that only three mutations within the GC box retain the ability to bind MIG1: G1 to C, C2 to T, and G5 to A. This result is consistent with current models for zinc finger-DNA binding, which assume that the sequence specificity is determined by base triplet recognition within the GC box. Surprisingly, we found that an AT-rich region 5' to the GC box also is important for MIG1 binding. This AT box is present in all natural MIG1 sites, and it is protected by MIG1 in DNase I footprints. However, the AT box differs from the GC box in that no single base within it is essential for binding. Instead, the AT-rich nature of this sequence seems to be crucial. The fact that AT-rich sequences are known to increase DNA flexibility prompted us to test whether MIG1 bends DNA. We found that binding of MIG1 is associated with bending within the AT box. We conclude that DNA binding by a simple zinc finger protein such as MIG1 can involve both recognition of the GC box and flanking sequence preferences that may reflect local DNA bendability.

scholarly journals Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1.

1994 ◽  
Vol 14 (3) ◽  
pp. 1979-1985 ◽  
Author(s):  
M Lundin ◽  
J O Nehlin ◽  
H Ronne
Keyword(s):  
Dna Binding ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Wilms Tumor ◽  
Glucose Repression ◽  
Saturation Mutagenesis ◽  
Sequence Specificity ◽  
Rich Region ◽  
Finger Protein ◽  
Gc Box

MIG1 is a zinc finger protein that mediates glucose repression in the yeast Saccharomyces cerevisiae. MIG1 is related to the mammalian Krox/Egr, Wilms' tumor, and Sp1 finger proteins. It has two fingers and binds to a GCGGGG motif that resembles the GC boxes recognized by these mammalian proteins. We have performed a complete saturation mutagenesis of a natural MIG1 site in order to elucidate its binding specificity. We found that only three mutations within the GC box retain the ability to bind MIG1: G1 to C, C2 to T, and G5 to A. This result is consistent with current models for zinc finger-DNA binding, which assume that the sequence specificity is determined by base triplet recognition within the GC box. Surprisingly, we found that an AT-rich region 5' to the GC box also is important for MIG1 binding. This AT box is present in all natural MIG1 sites, and it is protected by MIG1 in DNase I footprints. However, the AT box differs from the GC box in that no single base within it is essential for binding. Instead, the AT-rich nature of this sequence seems to be crucial. The fact that AT-rich sequences are known to increase DNA flexibility prompted us to test whether MIG1 bends DNA. We found that binding of MIG1 is associated with bending within the AT box. We conclude that DNA binding by a simple zinc finger protein such as MIG1 can involve both recognition of the GC box and flanking sequence preferences that may reflect local DNA bendability.

scholarly journals Two homologous zinc finger genes identified by multicopy suppression in a SNF1 protein kinase mutant of Saccharomyces cerevisiae.

1993 ◽  
Vol 13 (7) ◽  
pp. 3872-3881 ◽  
Author(s):  
F Estruch ◽  
M Carlson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Protein Kinase ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Wilms Tumor ◽  
Temperature Sensitive ◽  
Carbon Utilization ◽  
Functional Roles ◽  
Multicopy Suppressor ◽  
Finger Protein

The MSN2 gene was selected as a multicopy suppressor in a temperature-sensitive SNF1 protein kinase mutant of Saccharomyces cerevisiae. MSN2 encodes a Cys2His2 zinc finger protein related to the yeast MIG1 repressor and to mammalian early growth response and Wilms' tumor zinc finger proteins. Deletion of MSN2 caused no phenotype. A second similar zinc finger gene, MSN4, was isolated, and deletion of both genes caused phenotypic defects related to carbon utilization. Overexpression of the zinc finger regions was deleterious to growth. LexA-MSN2 and LexA-MSN4 fusion proteins functioned as strong transcriptional activators when bound to DNA. Functional roles of this zinc finger protein family are discussed.

scholarly journals Transcriptional Regulation of the CLC-K1 Promoter by myc-Associated Zinc Finger Protein and Kidney-Enriched Krüppel-Like Factor, a Novel Zinc Finger Repressor

2000 ◽  
Vol 20 (19) ◽  
pp. 7319-7331 ◽  
Author(s):  
Shinichi Uchida ◽  
Yujiro Tanaka ◽  
Hiroshi Ito ◽  
Fumiko Saitoh-Ohara ◽  
Johji Inazawa ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Mesangial Cells ◽  
Specific Binding ◽  
Consensus Sequence ◽  
Interstitial Cells ◽  
Luciferase Reporter ◽  
Gene Promoters ◽  
Finger Protein

ABSTRACT The expression of CLC-K1 and CLC-K2, two kidney-specific CLC chloride channels, is transcriptionally regulated on a tissue-specific basis. Previous studies have shown that a GA element near their transcriptional start sites is important for basal and cell-specific activities of the CLC-K1 and CLC-K2 gene promoters. To identify the GA-binding proteins, the human kidney cDNA library was screened by a yeast one-hybrid system. A novel member of the Cys2-His2 zinc finger gene designated KKLF (for “kidney-enriched Krüppel-like factor”) and the previously isolated MAZ (for “myc-associated zinc finger protein”) were cloned. KKLF was found to be abundantly expressed in the liver, kidneys, heart, and skeletal muscle, and immunohistochemistry revealed the nuclear localization of KKLF protein in interstitial cells in heart and skeletal muscle, stellate cells, and fibroblasts in the liver. In the kidneys, KKLF protein was localized in interstitial cells, mesangial cells, and nephron segments, where CLC-K1 and CLC-K2 were not expressed. A gel mobility shift assay revealed sequence-specific binding of recombinant KKLF and MAZ proteins to the CLC-K1 GA element, and the fine-mutation assay clarified that the consensus sequence for the KKLF binding site was GGGGNGGNG. In a transient-transfection experiment, MAZ had a strong activating effect on transcription of the CLC-K1–luciferase reporter gene. On the other hand, KKLF coexpression with MAZ appeared to block the activating effect of MAZ. These results suggest that a novel set of zinc finger proteins may help regulate the strict tissue- and nephron segment-specific expression of the CLC-K1 and CLC-K2 channel genes through their GA cis element.

scholarly journals Structural Insights into c-Myc-interacting Zinc Finger Protein-1 (Miz-1) Delineate Domains Required for DNA Scanning and Sequence-specific Binding

2016 ◽  
Vol 292 (8) ◽  
pp. 3323-3340 ◽  
Author(s):  
Mikaël Bédard ◽  
Vincent Roy ◽  
Martin Montagne ◽  
Pierre Lavigne
Keyword(s):  
Dna Binding ◽  
Zinc Finger ◽  
Dna Sequences ◽  
Zinc Finger Protein ◽  
Specific Binding ◽  
Consensus Sequence ◽  
Compact Structure ◽  
General Role ◽  
Finger Protein

c-Myc-interacting zinc finger protein-1 (Miz-1) is a poly-Cys2His2 zinc finger (ZF) transcriptional regulator of many cell cycle genes. A Miz-1 DNA sequence consensus has recently been identified and has also unveiled Miz-1 functions in other cellular processes, underscoring its importance in the cell. Miz-1 contains 13 ZFs, but it is unknown why Miz-1 has so many ZFs and whether they recognize and bind DNA sequences in a typical fashion. Here, we used NMR to deduce the role of Miz-1 ZFs 1–4 in detecting the Miz-1 consensus sequence and preventing nonspecific DNA binding. In the construct containing the first 4 ZFs, we observed that ZFs 3 and 4 form an unusual compact and stable structure that restricts their motions. Disruption of this compact structure by an electrostatically mismatched A86K mutation profoundly affected the DNA binding properties of the WT construct. On the one hand, Miz1–4WT was found to bind the Miz-1 DNA consensus sequence weakly and through ZFs 1–3 only. On the other hand, the four ZFs in the structurally destabilized Miz1–4A86K mutant bound to the DNA consensus with a 30-fold increase in affinity (100 nm). The formation of such a thermodynamically stable but nonspecific complex is expected to slow down the rate of DNA scanning by Miz-1 during the search for its consensus sequence. Interestingly, we found that the motif stabilizing the compact structure between ZFs 3 and 4 is conserved and enriched in other long poly-ZF proteins. As discussed in detail, our findings support a general role of compact inter-ZF structures in minimizing the formation of off-target DNA complexes.

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