• Molecules and Cells
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• 제23권3호
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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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• 제25권4호
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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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• 제34권3호
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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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• 제35권3호
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• pp.177-183
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• 2008

본 연구는 동진벼 유래의 Ac/Ds 삽입변이집단의 GUS 분석을 통하여 뿌리 및 미숙종자에서 강하게 GUS가 발현한 개체를 선발하여 FST(flanking sequence tag) 분석 한 결과 Ds 전이 인자가 3번 염색체 zinc finger RING-H2 관련 Oszinc626 유전자의 첫 번째 exon 부위에 single copy로 삽입되어 있었으며, 선발변이체는 뿌리 및 종자 발달이 정상인 동진벼에 비해 매우 낮은 것으로 나타났다. Oszinc626 유전자는 RING-H2 type(C3-H2-C3)으로 $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$ 배열이 C-terminus 가장 말단에 위치하며, 49 kDa의 분자량을 가지고 있다. 또한 Southern blot 분석에서 Oszinc626 유전자는 벼 게놈상에 single copy로 존재하였다. RT-PCR을 통한 돌연변이 유전자의 발현분석 결과 250 mM의 염과, $4^{\circ}C$ 저온등과 같은abiotic stress에 의해 발현이 증가함을 보였고, 호르몬처리에 있어서 ABA와 IAA의 식물호르몬을 처리했을 경우 24시간까지 계속해서 발현양이 증가하는 것을 보이는 반면, 2,4-D 처리의 경우 30분 후에 발현이 일시적으로 증가되었으나 이후 발현이 급속히 감소한 것을 보였다. 벼의 조직 별 발현 검정에서 미성숙한 종자, 뿌리 분열조직 및 신초 등 주로 생장점 부위에서 강하게 발현되는 것을 보임에 따라 Oszinc626 유전자의 경우 식물의 생장에 관여하는 주동 유전자의 하나로 판단된다.

• BMB Reports
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• 제40권4호
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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.

• 한국환경농학회지
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• 제27권4호
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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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• 제26권2호
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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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• 제53권3호
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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.

• 미생물학회지
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• 제32권2호
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• pp.102-108
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• 1994

ABF1(Autonomously replicating sequence Binding Factor 1)은 효모 genome에서 $RTCRYN_5ACG$의 염기 서열을 가지고 있는 promoter, mating-type silencer, ARS에 결합하는 DNA binding 단백질이다. E. coli 에서 ABF1 유전자를 발현하기 위하여, ABF1 유전자를 pMAL-c2 벡터에 cloning하였다.(pMAHW). pMAHW를 E. coli에 형질전환하여, ABF1 융합단백질을 발현시키고, amylose resin affinity chromatography에 의하여 분리하였다. Factor Xa protease를 이용하여 분리된 융합단백질로부터 maltose binding protein을 잘라낸 후에 gel retardation analysis 방법으로 분리된 ABF1이 ARS1에 결합하는 능력을 지니고 있음을 확인하였다. DNA 결합에 관련된 부위를 찾기 위하여, 비전형적인 zinc finger motif가 위치하는 자리에서 pMAHW의 ABF1 유전자에 His-61을 다른 아미노산으로 치환하였다. DNA binding 부위로 추정되는 ABF1 단백질의 중간지역에 Leu-353, Leu-360를 다른 아미노산으로 치환하였다. Site-specific mutagenesis 를 통해 만들어진 mutant를 gel retardation analysis와 complementation test를 통해서 비전형적인 zinc finger motif이외에 다른 DNA binding motif가 있는 것을 알 수 있었다.

• 미생물학회지
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• 제42권4호
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• pp.246-251
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• 2006

사상성 진균인 Aspergillus nidulans에서 유성분화초기단계, 또는 유성분화유도를 위한 세포내 조건 형성과정에 관여할 것으로 예상되는 유전자를 탐색하였다. 선행연구결과를 통해 유성분화를 전혀 하지 못하는 NSD (never in sexual development) 돌연변이주가 분리되어 nsdA, nsdB, nsdC, 그리고 nsdD의 4상 보군으로 동정된 바 있다. 본 연구에서는 이들 유전자 중 nsdC 유전자를 분리하고자 A. nidulans AMAl-Not I Genomic DNA library로 nsdC6 돌연변이균주를 형질전환하여 야생형처럼 유성분화를 할 수 있는 형질전환체를 분리하고 이들로부터 약 10 kb genomic DNA가 삽입된 library DNA를 분리하였다. Genomic priming system (GPS)을 이용하여 nsdC6 돌연변이를 상보하는 유전자의 부분 서열을 확보한 후 전체 DNA 염기서열을 결정하였다. 유전자분석 결과 nsdC는 intron 없이 1,929염기(643개의 아미노산)로 구성된 Open reading frame (ORF)를 가지며, 약 1kb 정도의 비교적 긴 5'-UTR 부위에 2개의 intron을 가지고 있음이 확인되었다. 또한 NsdC polypeptide의 중앙에 $C_2H_2C_2H_2C_2HC$ 형의 zinc finger DNA binding domain과 C 말단 부위에 coiled-coil domain이 존재하였다. nsdC6 돌연변이는ORF의 407 bp와 408 bp사이에 엽기 T가 삽입되어 frameshift가 일어난 것으로 밝혀졌다. 따라서 nsdC6 돌연변이균주는 단지 139개 아미노산만 갖고 있는 결실 단백질이 생산됨을 알 수 있었다.