• Molecules and Cells
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• v.23 no.3
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• pp.316-322
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• 2007

Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

• The Plant Pathology Journal
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• v.25 no.4
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• pp.381-388
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• 2009

On screening pathogen-resistant soybean, we identified a WRKY type transcription factor named a Glycine max WRKY1 (GmWRKY1). Expression of GmWRKY1 gene was induced in the soybean sprout by Pseudomonas infection. The GmWRKY1 was expressed in all of the tissues with high levels in stems, leaves and developing seeds. The protein Gm WRKY1 contains highly conserved two WRKY DNA-binding domains having two $C_2-H_2$ zinc-finger motif ($C-X_{4-5}-C-X_{22-23}-H-X-H$) in its N-terminal and C-terminal amino acid sequences. In electrophoresis mobility shift assay, the GmWRKY1 protein bound specifically to W-box elements in the promoters of defense related genes. These results demonstrated that GmWRKY1 is one of the soybean WRKY family genes and the plant-specific transcription factors for defense processes.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.167-172
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• 2007

A tomato zinc-finger protein gene, LeZFP1, encoding the Cys2/His2-type zinc-finger transcription factor was searched from cDNA microarray analysis of gene expression following induction of the overexpressed tomato transgenic plants showing resistance for pathogen and abiotic stresses. The full-length cDNA of LeZFP1 encoded a protein of 261 amino acid residues. Analysis of the deduced amino acid sequence of LeZFP1 revealed that it shares high sequence identity with pepper CAZFP1 (81% identity). We found that single copy of LeZFP1 gene is present in the tomato genome through southern blot analysis. The LeZFP1 transcripts were constitutively expressed in the tomato mature and young leaves, but were detectable weakly in the flower, stem and root. The LeZFP1 transcripts were significantly reduced in treated leaf tissues with NaCl and mannitol. The LeZFP1 gene was induced by oxidative stress especially. Our results indicated that LeZFP1 may play a role function involved in oxidative stress signaling pathways.

• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.177-183
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• 2008

Ac/Ds mutant lines of this study were transgenic rice plants, each of which harbored the maize transposable element Ds together with a GUS coding sequence under the control of a promoterless(Ds-GUS). We selected the mutants that were GUS expressed lines, because the GUS positive lines will be useful for identifying gene function in rice. One of these mutants was identified knock-out at Oszinc626(NP_001049991) gene, encoding a RING-H2 zinc-finger protein, by Ds insertion. In this mutant, while primary root development is normal, secondary root development from lateral root was very poor and seed development was incomplete compare with normal plant. RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, a few genes encoding RING zinc-finger proteins have been documented with visible effects on plant growth and development. The consensus of the RING-H2(C3-H2-C3 type) domain for this group of protein is $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$. Oszinc626 encodes a predicted protein product of 445 amino acids residues with a molecular mass of 49 kDa, with a RING-zinc-finger motif located at the extreme end of the C-terminus. RT-PCR analysis indicated that the expression of Oszinc626 gene was induced by IAA, cold, dehydration, high-salinity and abscisic acid, but not by 2,4-D, and the transcription of Oszinc626 gene accumulated primarily in rice immature seeds, root meristem and shoots. The gene accumulation patterns were corresponded with GUS expression.

• BMB Reports
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• v.40 no.4
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• pp.517-524
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• 2007

There were many different families of zinc finger proteins that contained multiple cysteine and/or histidine residues and used zinc to stabilize their folds. The classical C2H2 zinc finger proteins were the founding members of this superfamily and were among the most abundant proteins in eukaryotic genomes. C2H2 proteins typically contained several C2H2 fingers that made tandem contacts along the DNA. Here we reported a novel C2H2 type zinc finger gene, ZNF438, which encoded 828 amino acids that formed five zinc finger domains. Bioinformatics analysis revealed that the ZNF438 was mapped to human chromosome 10p11.2 and shared 62% identity with rat and mouse homologues. RT-PCR analysis indicated that it was ubiquitously expressed in 18 human adult tissues. With immunofluorescence assay, it was shown that the exogenous Flag-tagged ZNF438 was located in nucleus of COS-7 cells. To further explore the function of ZNF438, we examined the transcriptional activity of ZNF438 protein by transfecting recombinant pM-ZNF438 into mammalian cells. The subsequent analysis based on the duel luciferase assay system showed that ZNF438 was a transcriptional repressor.

• Korean Journal of Environmental Agriculture
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• v.27 no.4
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• pp.389-394
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• 2008

The ecophysiological changes occurring upon cold stress were studied using cold tolerant transgenic and wild-type tobacco plants. In a previous study, cold tolerance in tobacco was induced by the introduction of a gene encoding the zinc finger transcription factor, PIF1. Gas-exchange measurements including net photosynthesis and stomatal conductance were performed prior to, in the middle of, and after a cold-stress treatment of $1{\pm}2^{\circ}C$ for 96 h in each of the four seasons. In both transgenic and wild-type plants, gas-exchange parameters were severely decreased in the middle of the cold treatment, but had recovered after 2-3 h of adaptation in a greenhouse. Most t-test comparisons on gas-exchange measurements between the two plant types did not show statistical significance. Wild-type plants had slightly more water-soaked damage on the leaves than the transgenic plants. A light-response curve did not show any differences between the two plant types. However, the curve for assimilation-internal $CO_2$ in wild-type plants showed a much higher slope than that of the PIF1 transgenic plants. This means that the wild-type plant is more capable of regenerating Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and has greater electron transport capacity. In conclusion, cold-resistant transgenic tobacco plants demonstrated a better recovery of net photosynthesis and stomatal conductance after cold-stress treatment compared to wild-type plants, but the ecophysiological recoveries of the transgenic plants were not statistically significant.

• Molecules and Cells
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• v.26 no.2
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• pp.175-180
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• 2008

$I{\kappa}B$ kinase (IKK), the pivotal kinase in signal-dependent activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$), is composed of multiple protein components, including IKK ${\alpha}/{\beta}/{\gamma}$ core subunits. To investigate the regulation of the IKK complex, we immunoaffinity purified the IKK complex, and by MALDI-TOF mass spectrometry identified a splice variant of zinc finger protein 268 (ZNF268) as a novel IKKinteracting protein. Both the full-length and the spliced form of the ZNF268 protein were detected in a variety of mammalian tissues and cell lines. The genes were cloned and expressed by in vitro transcription/translation. Several deletion derivatives, such as KRAB domain (KRAB) on its own, the KRAB/spacer/4-zinc fingers (zF4), and the spacer/4-zinc fingers (zS4), were ectopically expressed in mammalian cells and exhibited had different subcellular locations. The KRAB-containing mutants were restricted to the nucleus, while zS4 was localized in the cytosol. TNF-${\alpha}$-induced NF-${\kappa}B$ activation was examined using these mutants and only zS4 was found to stimulate activation. Collectively, the results indicate that a spliced form of ZNF268 lacking the KRAB domain is located in the cytosol, where it seems to play a role in TNF-${\alpha}$-induced NF-${\kappa}B$ activation by interacting with the IKK complex.

• BMB Reports
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• v.53 no.3
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• pp.160-165
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• 2020

The root meristem of Arabidopsis thaliana is protected by the root cap, the size of which is tightly regulated by the balance between the formative cell divisions and the dispersal of the outermost cells. We isolated an enhancer-tagged dominant mutant displaying the short and twisted root by the overexpression of ZINC-FINGER OF ARABIDOPSIS THALIANA1 (ZAT1) encoding an EAR motif-containing zinc-finger protein. The growth inhibition by ZAT1 was shared by ZAT4 and ZAT9, the ZAT1 homologues. The ZAT1 promoter was specifically active in the outermost cells of the root cap, in which ZAT1-GFP was localized when expressed by the ZAT1 promoter. The outermost cell-specific expression pattern of ZAT1 was not altered in the sombrero (smb) or smb bearskin1 (brn1) brn2 accumulating additional root-cap layers. In contrast, ZAT4-GFP and ZAT9-GFP fusion proteins were distributed to the inner root-cap cells in addition to the outermost cells where ZAT4 and ZAT9 promoters were active. Overexpression of ZAT1 induced the ectopic expression of PUTATIVE ASPARTIC PROTEASE3 involved in the programmed cell death. The EAR motif was essential for the growth inhibition by ZAT1. These results suggest that the three related ZATs might regulate the maturation of the outermost cells of the root cap.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.102-108
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• 1994

Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

• Korean Journal of Microbiology
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• v.42 no.4
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• pp.246-251
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• 2006

A lot of mutants which cannot initiate sexual development were screened and several loci including nsdA, nsdB, nsdC, and nsdD were identified in homothallic ascomycetes Aspergillus nidulans. The NSD206, which has nsdC6 allele, showed typical phenotype of NSD (Never in sexual development) mutants. The nsdC gene was cloned by transforming NSDP697 ($nsdC^-$, $pryG^-$) with AMA1-NotI genomic library. The transforming library DNA recovered from several transformants showing wild phenotype carried about 10 kb genomic DNA insert. The DNA sequence of nsdC was analysed using GPS (Genome priming system). The nsdC gene has an open reading frame (ORF) of 1,929 bp encoding a putative polypeptide of 643 amino acids. The NsdC carries $C_2H_2C_2H_2C_2HC$ type zinc finger DNA binding domains in the middle of the polypeptide. A coiled-coil domain at its C terminus were also found. In nsdC6 allele, a single T insertion was occurred between 407-408 bp leading to the frameshift mutation and early termination of translation producing the truncated protein which has only 139 amino acids.