Cloning the cDNA for a new human zinc finger protein defines a group of closely related krüppel like transcription factors. (1998 Nov)
cloning the cDNA for a new human zinc finger protein defines a group of closely related krüppel like transcription factors . We have identified a novel zinc finger protein that has been named ubiquitous krüppel like factor ( UKLF ) based on structural considerations and the pattern of gene expression . UKLF was isolated by the polymerase chain reaction approach using degenerate oligonucleotides corresponding to the DNA binding domain of erythroid krüppel like factor ( EKLF ) and cDNA prepared from human vascular endothelial cells . The carboxyl terminal portion of UKLF contains three zinc fingers of the Cys2 His2 type and binds in vitro to the caccc motif of the beta globin promoter and to the Sp1 recognition sequence . The amino terminal portion of UKLF consists of a hydrophobic region rich in serines and a negatively charged segment with several glutamic acid residues . The first 47 amino acids of the acidic region are nearly identical to the amino terminal portion of another krüppel like factor , the so called core promoter binding protein ( CPBP ) or Zf9 . Like CPBP / Zf9 , UKLF can function as a transcription activator in co transfection assays . however , this activity is lost when the highly conserved amino terminal segment is deleted . these findings indicate that UKLF and CPBP / Zf9 represent a distinct subgroup of closely related krüppel like activators of transcription . mapping of the UKLF gene to chromosome 2 suggested that UKLF and CPBP …

Zinc finger transcription factors designed for bispecific coregulation of erbb2 and erbb3 receptors : insights into ErbB receptor biology. (2005 Oct)
Zinc finger transcription factors designed for bispecific coregulation of erbb2 and erbb3 receptors : insights into ErbB receptor biology . signaling through the ErbB family of tyrosine kinase receptors in normal and cancer derived cell lines contributes to cell growth and differentiation . In this work , we altered the levels of erbb2 and erbb3 receptors , individually and in combination , by using 6 finger and 12 finger synthetic zinc finger protein artificial transcription factors ( ATFs ) in an epidermoid squamous cell carcinoma line , A431 . We successfully designed 12 finger ATFs capable of coregulating erbb3 and ICAM 1 or erbb2 and erbb3 . With ATFs , the effects of changes in erbb2 and erbb3 receptor levels were evaluated by using cell proliferation , cell migration , and cell signaling assays . Cell proliferation was increased when erbb2 and erbb3 were both overexpressed . Cell migration on collagen was decreased when erbb2 was down regulated , yet migration on laminin was significantly increased with erbb3 overexpression . erbb2 and erbb3 overexpression also stimulated the phosphatidylinositol 3 kinase and mitogen activated protein kinase pathways . Our ATF approach has elucidated differences in ErbB receptor mediated proliferation , migration , and intracellular signaling that cannot be explained merely by the presence or absence of particular ErbB receptors and emphasizes the dynamic nature of the ErbB signaling system . The transcription factor approach developed here provides a gene economical route to the regulation of multiple genes and may be important …

Influence of parenteral zinc and actinomycin D on tissue zinc uptake and the synthesis of a zinc binding protein. (1975 Aug)
influence of parenteral zinc and actinomycin D on tissue zinc uptake and the synthesis of a zinc binding protein . parenterally administered zinc markedly increased the incorporation of 14 C cystine and 65 Zn into a low molecular weight zinc binding protein ( ZnBP ) isolated from liver cytoplasm of rats fed an adequate amount of zinc . This zinc load significantly increased the zinc content in the liver . The increase in hepatic zinc content was inhibited by actinomycin D indicating that DNA dependent RNA synthesis is required for zinc uptake into liver . antinomycin D also produced a concomitant decrease in ZnBP synthesis indicating that this protein may be involved in the uptake mechanism in cells . Zinc repletion also stimulated the synthesis of hepatic ZnBP in zinc deficient rats . This stimulation was also prevented by prior administration of actinomycin D . A similar effect was observed in the intestinal mucosal cells . The data collectively indicate that the control of the synthesis of ZnBP which occurs at the transcriptional level of protein synthesis is responsive to zinc status and thus may have a function in zinc metabolism .

The ribonuclease activity of the two synthetic polypeptides having zinc finger sequence. (1993 Oct)
The ribonuclease activity of the two synthetic polypeptides having zinc finger sequence .

Identification of a novel krueppel related zinc finger gene ( znf184 ) mapping to 6p21. (1997 May)
identification of a novel krueppel related zinc finger gene ( znf184 ) mapping to 6p21 . 3 . cDNA selection and exon trapping were performed on cosmids mapping to a region 3 Mb distal to HLA A . analysis of resulting fragments indicated the presence of two zinc finger transcripts , and one of these was used to isolate a partial cDNA ( znf184 ) from a placental library . The second transcript contained additional sequence of the 5 end of the gene , extending the sequence to 2678 bp . sequence analysis indicates that znf184 is a classical krueppel zinc finger with 19 highly conserved zinc finger motifs at the C terminus and a krueppel associated box at the N terminus of the protein . This gene encodes a 3 . 2 kb transcript that is highly expressed in testis and expressed at a moderate to low level in all other tissues tested . This zinc finger gene maps to a region approximately 200 kb distal to the microsatellite marker d6s105 and approximately 300 kb proximal to d6s1260 .

The biology of the mammalian krüppel like family of transcription factors. (2001 Jan)
The biology of the mammalian krüppel like family of transcription factors . recent advances in molecular cloning have led to the identification of a large number of mammalian zinc finger containing transcription factors that exhibit homology to the drosophila melanogaster protein , krüppel . although the amino acid sequences in the zinc finger domains of these krüppel like factors ( KLFs ) are closely related to one another , the regions outside the zinc fingers of the proteins are usually unique . KLFs display seemingly different and broad biological properties with each functioning as an activator of transcription , a repressor or both . This review article provides a current phylogenetic classification of the identified KLFs to date . More importantly , the currently known biological activities of the KLFs in regulating transcription , cell proliferation , differentiation and development are summarized and compared . further characterization of this interesting protein family should provide additional insights into the their respective regulatory role in various important biological processes .

Identification , cloning and sequence of the aspergillus niger areA wide domain regulatory gene controlling nitrogen utilisation. (1998 Apr)
identification , cloning and sequence of the aspergillus niger areA wide domain regulatory gene controlling nitrogen utilisation . The gene encoding the positive acting regulator of nitrogen metabolite repression ( AREA ) has been cloned and characterised from the industrially important filamentous fungus aspergillus niger . The deduced amino acid sequence has an overall level of identity with its homologues from other fungal species which varies between 32 and 72 . This gene ( areanig ) complements the A . nidulans arear 18 loss of function mutation . sequences upstream of the structural gene contain several putative GATA type zinc finger protein binding motifs . northern analysis indicates the synthesis of multiple transcripts , the major species being approximately 2 . 95 kb and 3 . 1 kb . maximal expression of areanig is observed under conditions of nitrogen starvation and is mainly due to an increase in the level of the shorter transcript .

Transcriptional activation by the papillomavirus E6 zinc finger oncoprotein. (1990 Jul)
transcriptional activation by the papillomavirus E6 zinc finger oncoprotein . The introduction of the bovine ( BPV ) or human papillomavirus E6 gene into susceptible cells can result in their transformation , but there are few clues to the mechanism of action of the E6 gene . The characteristic features of E6 proteins are their small size ( approximately 150 amino acids ) and the potential to form two large zinc fingers . To determine if E6 can function as a transcription factor , the BPV E6 gene was fused to the sequence specific DNA binding peptide encoded by the BPV E2 gene . This chimeric E6 E2 protein trans activated promoters that incorporated E2 binding elements in both rodent cells and saccharomyces cerevisiae . In the absence of E6 E2 localization to the target promoter , trans activation did not occur . alteration of the cysteine residues at the base of each finger abrogated the transcriptional activity of the E6 E2 hybrids . these data demonstrated that the BPV E6 gene encodes a transcription activation domain and imply that a specific structure of the protein , most likely the zinc fingers , is critical for this function . since these cysteine mutants are also transformation defective , E6 transcriptional functions may be required for its oncogenic activity .

Molecular cloning of a zinc finger protein which binds to the heptamer of the signal sequence for V ( D … (1994 Jan)
molecular cloning of a zinc finger protein which binds to the heptamer of the signal sequence for V ( D ) J recombination . The somatic V ( D ) J recombination for the assembly of the Ig and TCR genes is mediated by the recombination signal sequences ( Rss ) and the V ( D ) J recombinase . A cDNA clone was isolated from a lambda gt11 expression library made from mouse thymocyte poly ( A ) RNA , using the Rss as a ligand . The deduced amino acid sequence of the putative protein , designated recognition component ( Rc ) , reveals a pair of Cys2 His2 zinc fingers followed by a Glu and Asp rich acidic domain . In addition , there are five copies of the Ser / Thr Pro X Arg / Lys sequence , which are putative DNA binding units . The zinc finger acidic domain structures present in Rc are also found in several enhancer binding proteins , such as those for the kappa B motif of the Ig kappa light chain enhancer or related sequences . bacterial fusion proteins for Rc bind preferentially to the Rss heptamer and to the kappa B motif . The dual affinities of Rc for the Rss heptamer and the kappa B motif suggest a possible link between Ig transcription and somatic recombination . The formation of multiple gel shifted DNA protein complexes for Rc and its DNA ligand suggests that these complexes tend to …

Genomic organisation and characterisation of the neural sex determination gene fruitless ( fru ) in the hawaiian species drosophila heteroneura. (2000 May)
genomic organisation and characterisation of the neural sex determination gene fruitless ( fru ) in the hawaiian species drosophila heteroneura . there are several mechanisms for the determination of sex . sexual behaviour is part of the sex determination cascade , and in drosophila melanogaster male courtship is controlled in part by the fruitless gene . As part of a study of sexual behaviour in hawaiian drosophila , we have cloned the neural sex determination gene fru from the hawaiian picture wing species drosophila heteroneura . The fru gene has at least seven exons covering a region of 18kb and encodes three transcripts , fruA , fruB and fruC . Each transcript encodes a single ORF of 841 , 678 and 691aa , respectively . The FRUA and FRUB proteins have a BTB protein protein binding domain and two zinc finger like domains and are well conserved with the D . melanogaster proteins . The FRUC protein has a BTB domain but no zinc finger like domains . The fru gene is expressed in 1 7 day old adult males as a 5 . 1kb transcript . This transcript is not seen in adult females , so the fru gene has a different pattern of sex differential expression in the hawaiian drosophila compared with D . melanogaster .

The arabidopsis jagged gene encodes a zinc finger protein that promotes leaf tissue development. (2004 Feb)
The arabidopsis jagged gene encodes a zinc finger protein that promotes leaf tissue development . important goals in understanding leaf development are to identify genes involved in pattern specification , and also genes that translate this information into cell types and tissue structure . Loss of function mutations at the jagged ( JAG ) locus result in arabidopsis plants with abnormally shaped lateral organs including serrated leaves , narrow floral organs , and petals that contain fewer but more elongate cells . jag mutations also suppress bract formation in leafy , apetala1 and apetala2 mutant backgrounds . The JAG gene was identified by map based cloning to be a member of the zinc finger family of plant transcription factors and encodes a protein similar in structure to superman with a single C ( 2 ) H ( 2 ) type zinc finger , a proline rich motif and a short leucine rich repressor motif . JAG mRNA is localized to lateral organ primordia throughout the plant but is not found in the shoot apical meristem . misexpression of JAG results in leaf fusion and the development of ectopic leaf like outgrowth from both vegetative and floral tissues . Thus , JAG is necessary for proper lateral organ shape and is sufficient to induce the proliferation of lateral organ tissue .

The RIM / NIM family of neuronal C2 domain proteins. (2000 Aug)
The RIM / NIM family of neuronal C2 domain proteins . interactions with Rab3 and a new class of Src homology 3 domain proteins . RIM1 is a putative effector protein for rab3s , synaptic GTP binding proteins . RIM1 is localized close to the active zone at the synapse , where it interacts in a GTP dependent manner with Rab3 located on synaptic vesicles . We now describe a second RIM protein , called RIM2 , that is highly homologous to RIM1 and also expressed primarily in brain . Like RIM1 , RIM2 contains an N terminal zinc finger domain that binds to Rab3 as a function of GTP , a central PDZ domain , and two C terminal C ( 2 ) domains that are separated by long alternatively spliced sequences . unexpectedly , the 3 end of the RIM2 gene produces an independent mRNA that encodes a smaller protein referred as NIM2 . NIM2 is composed of a unique N terminal sequence followed by the C terminal part of RIM2 . Data bank searches identified a third RIM / NIM related gene , which encodes a NIM isoform referred to as NIM3 ; no RIM transcript from this gene was detected . To test if NIMs , like RIMs , may function in secretion , we investigated the effect of NIM3 on calcium triggered exocytosis in PC12 cells . NIM3 induced a dramatic increase in calcium evoked exocytosis ( 50 ) , with no significant effect on …

The zinc finger of the CSN associated deubiquitinating enzyme usp15 is essential to rescue the E3 ligase Rbx1. (2005 Jul)
The zinc finger of the CSN associated deubiquitinating enzyme usp15 is essential to rescue the E3 ligase Rbx1 . The COP9 signalosome ( CSN ) is a conserved protein complex found in all eukaryotic cells and involved in the regulation of the ubiquitin ( Ub ) / 26S proteasome system . It binds numerous proteins , including the Ub E3 ligases and the deubiquitinating enzyme ubp12p , the S . pombe ortholog of human usp15 . We found that usp15 copurified with the human CSN complex . isolated CSN complex exhibited protease activity that deubiquitinated poly Ub substrates and was completely inhibited by o phenanthroline ( OPT ) , a metal chelating agent . surprisingly , the recombinant usp15 was also not able to cleave isopeptide bonds of poly Ub chains in presence of OPT . detailed analysis of USP sequences led to the discovery of a novel zinc ( Zn ) finger in usp15 and related USPs . mutation of a single conserved cysteine residue in the predicted Zn binding motif resulted in the loss of usp15 capability to degrade poly Ub substrates , indicating that the Zn finger is essential for the cleavage of poly Ub chains . moreover , pulldown experiments demonstrated diminished binding of tetra Ub to mutated usp15 . cotransfection of usp15 and the Ub ligase Rbx1 revealed that the wild type deubiquitinating enzyme , but not the usp15 mutant with a defective Zn finger , stabilized Rbx1 toward the Ub system , most likely …

Cloning , genomic organization and promoter activity of the mouse zinc finger protein gene ZF 12 The human zinc finger … (2003 Jun)
cloning , genomic organization and promoter activity of the mouse zinc finger protein gene ZF 12 The human zinc finger protein znf191 is a krüppel like transcription factor , which may be relevant to many diseases such as neuropsychiatric , cardiovascular and liver caner diseases . To elucidate the function of znf191 by gene targeting , it is necessary to clone and characterize of the homologous gene in model organisms ( mice ) . The mouse homologous gene ( ZF 12 ) was cloned and sequenced for the first time , the genbank accession number is ay052495 . It contains four exons and three introns ; all intronic splice sites exhibited consensus GT / AG sequences . The single nucleotide polymorphisms ( SNPs ) in exon 2 and the alternative length of 3 untranslated region ( 3 UTR ) have been found . The linkage of the ZF 12 gene and the zinc finger protein gene Zfp 35 has been found , so the ZF 12 gene can be localized to B3 to C or beside of chromosome 18 . We assessed approximately 1 . 2 kb of 5 flanking region of the ZF 12 gene for basal promoter activity . A series of deletion mutants of 5 flanking region linked to the luciferase gene was constructed . basal level expression of these constructs was tested in COS 7 cells , nih3t3 cells and HeLa cells . By measuring luciferase activity , which was transiently expressed in the transfected cells …

Recruitment of CD40 and tumor necrosis factor receptor associated factors 2 and 3 to membrane microdomains during CD40 signaling. (2000 Jun)
recruitment of CD40 and tumor necrosis factor receptor associated factors 2 and 3 to membrane microdomains during CD40 signaling . signals delivered to antigen presenting cells through CD40 are critical for the activation of immune responses . intracellular tumor necrosis factor ( TNF ) receptor associated factors ( trafs ) are key elements of the signal transduction pathways of many TNF receptor family members , including CD40 . We show for the first time that engagement of CD40 in intact B cells induces the rapid translocation of traf2 from the cytoplasm to the plasma membrane . We found that CD40 engagement also results in its recruitment , together with traf2 and traf3 , to membrane microdomains , regions of the plasma membrane enriched in signaling molecules such as the Src family kinases . using a membrane permeable chelator of zinc or a mutant traf2 molecule , we show that the putative zinc binding domains of trafs contribute to their recruitment to microdomains and to the downstream activation of c Jun N terminal kinase . We suggest that the zinc RING and zinc finger domains of trafs are required for communication between CD40 and microdomain associated signaling molecules and may serve a similar role in the signal transduction pathways of other TNF receptor family members .

Cloning and characterization of a sulphite resistance gene of saccharomyces cerevisiae. (1995 Jan)
cloning and characterization of a sulphite resistance gene of saccharomyces cerevisiae . In this paper we describe the cloning and sequencing of the gene ( SUL1 ) responsible for sulphite resistance in a saccharomyces cerevisiae mutant ( casalone et al . , 1992 ) . The deduced amino acid sequence predicted that the gene codes for a zinc finger protein with five fingers . comparison of wild type and mutant gene sequences demonstrated that the mutation event was a transversion from C to G ; as a consequence of the mutation a histidine substituted an aspartic acid , affecting directly the fourth finger structure . The SUL1 gene sequence corresponds to that of FZF1 gene ( breitwieser et al . , 1993 ) to which no function was attributed .

Activation of the zinc finger encoding gene krox 20 in adult rat brain : comparison with zif268. (1992 Jun)
activation of the zinc finger encoding gene krox 20 in adult rat brain : comparison with zif268 . zif268 and krox 20 are transcription regulatory factors that contain highly homologous zinc finger DNA binding domains . recent studies have demonstrated that zif268 expression is rapidly regulated in brain by neuronal stimulation . We now report that , like zif268 , krox 20 is rapidly and transiently activated by electroconvulsive shock treatment ( ECT ) , D1 dopamine receptor activation , and opiate withdrawal . these studies indicate that , as found for the leucine zipper family of transcription factors , multiple members of the zinc finger family of transcription factors are induced by neuronal stimulation .

Why zinc fingers prefer zinc : ligand field symmetry and the hidden thermodynamics of metal ion selectivity. (2004 Nov)
Why zinc fingers prefer zinc : ligand field symmetry and the hidden thermodynamics of metal ion selectivity . The zinc finger , a motif of protein nucleic acid recognition broadly conserved among eukaryotes , is a globular minidomain containing a tetrahedral metal binding site . preferential coordination of Zn ( 2 ) ( relative to Co ( 2 ) ) is proposed to reflect differences in ligand field stabilization energies ( lfses ) due to complete or incomplete occupancy of d orbitals . LFSE predicts that the preference for Zn ( 2 ) should be purely enthalpic in accord with calorimetric studies of a high affinity consensus peptide ( CP 1 ; blasie , C . A . , and Berg , J . ( 2002 ) biochemistry 41 , 15068 73 ) . despite its elegance , the general predominance of LFSE is unclear as ( i ) the magnitude by which CP 1 prefers Zn ( 2 ) is greater than that expected and ( ii ) the analogous metal ion selectivity of a zinc metalloenzyme ( carbonic anhydrase ) is driven by changes in entropy rather than enthalpy . because CP 1 was designed to optimize zinc binding , we have investigated the NMR structure and metal ion selectivity of a natural finger of lower stability derived from human tumor suppressor protein WT1 . raman spectroscopy suggests that the structure of the WT1 domain is unaffected by interchange of Zn ( 2 ) and Co ( 2 …

Isolation and characterisation of a diverse family of arabidopsis two and three fingered C2H2 zinc finger protein genes and cdnas. (1997 Apr)
isolation and characterisation of a diverse family of arabidopsis two and three fingered C2H2 zinc finger protein genes and cdnas . In animal systems the C2H2 zinc finger protein ( ZFP ) gene family is the largest group of regulatory proteins and its members have a wide and important role in growth and development . It is likely that this family of ZFP transcription factors will also be important in plants . We have used a PCR approach employing highly degenerate oligonucleotide primers to isolate several arabidopsis genomic and cDNA clones encoding potential ZFPs . In addition we have used the sequence information from these clones to identify two ESTs as members of this family . Five two fingered and one three fingered ZFPs have been identified . outside of the zinc finger regions , there is considerable sequence diversity , including the sequence and length of the interfinger region ; the expression pattern of each gene is different . This is the first report of the isolation of a three fingered plant C2H2 ZFP gene and we discuss its possible evolutionary origin from two fingered ZFP genes .

Synthetic zinc finger peptides : old and novel applications. (2004 Jul)
synthetic zinc finger peptides : old and novel applications . In the last decade , the efforts in clarifying the interaction between zinc finger proteins and DNA targets strongly stimulated the creativity of scientists in the field of protein engineering . In particular , the versatility and the modularity of zinc finger ( ZF ) motives make these domains optimal building blocks for generating artificial zinc finger peptides ( ZFPs ) . ZFPs can act as transcription modulators potentially able to control the expression of any desired gene , when fused to an appropriate effector domain . artificial ZFPs open the possibility to re program the expression of specific genes at will and can represent a powerful tool in basic science , biotechnology and gene therapy . In this review we will focus on old , novel and possible future applications of artificial ZFPs .