RESUMO
We developed a rapid method to determine DNA-binding sites for putative DNA-binding proteins. This procedure has been successfully used to define a specific consensus site for the human ZNF35 zinc finger gene. ZNF35 encodes a 58-kDA polypeptide containing 11 consecutive finger motifs located at the amino terminus, and an acidic domain located at the carboxy terminus. These features suggest that ZNF35 is a site-specific DNA-binding protein involved in the regulation of gene expression. We have expressed the ZNF35 protein from E. coli and have employed a Southwestern-polymerase chain reaction method using random oligonucleotides to identify its high-affinity binding site. The core sequence for the ZNF35 protein-binding site is 5'-C/GC/GAAG/TA-3'.
Assuntos
Proteínas de Ligação a DNA/genética , Dedos de Zinco/genética , Sequência de Bases , Sítios de Ligação , Clonagem Molecular , Proteínas de Ligação a DNA/isolamento & purificação , Proteínas de Ligação a DNA/metabolismo , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Humanos , Fatores de Transcrição Kruppel-Like , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos/síntese química , Oligodesoxirribonucleotídeos/metabolismo , Plasmídeos , Reação em Cadeia da Polimerase/métodos , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
The evolutionarily conserved Krüppel-associated box (KRAB) is present in the N-terminal regions of more than one-third of all Krüppel-class zinc finger proteins. Recent experiments have demonstrated that the KRAB-A domain tethered to a promoter DNA by connecting to heterologous DNA-binding protein domain or targeted to a promoter-proximal RNA sequence acts as a transcriptional silencing of RNA polymerase II promoters. Here we show that expression of KRAB domain suppresses in vivo the activating function of various defined activating transcription factors, and we demonstrate that the KRAB domain specifically silences the activity of promoters whose initiation is dependent on the presence of a TATA box. Promoters whose accurate transcription initiation is directed by a pyrimidine-rich initiator element, however, are relatively unaffected. We also report in vitro transcription experiments indicating that the KRAB domain is able to repress both activated and basal promoter activity. Thus, the KRAB domain appears to repress the activity of certain promoters through direct communication with TATA box-dependent basal transcription machinery.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Regiões Promotoras Genéticas , RNA Polimerase II/biossíntese , RNA Polimerase II/genética , Fatores de Transcrição/metabolismo , Transcrição Gênica , Sequência de Bases , Cloranfenicol O-Acetiltransferase/análise , Cloranfenicol O-Acetiltransferase/biossíntese , Sequência Conservada , Repressão Enzimática , Repetição Terminal Longa de HIV , Células HeLa , Humanos , Fatores de Transcrição Kruppel-Like , Dados de Sequência Molecular , Oligodesoxirribonucleotídeos , Plasmídeos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Proteínas Repressoras/metabolismo , Mapeamento por Restrição , TATA Box , TransfecçãoRESUMO
Sub-families of related zinc finger protein genes have been defined on the basis of evolutionarily conserved structural features found outside the C2-H2 finger repeats. Such elements include the FAX domain found in a large number of Xenopus ZFPs, the evolutionarily conserved KRAB (Krüppel-associated box) and the ZiN (zinc finger N-terminal) domains. Here we describe a new evolutionarily conserved motif within zinc finger proteins which we have named the leucine rich region (LeR). Since conserved modules in regulatory proteins may specify properties relevant to their action we have determined the functional capabilities of LeR and the KRAB domains in the regulation of gene transcription by fusing relevant regions to a heterologous DNA-binding domain (GAL4 DNA-binding domain). We found that the KRAB-A domain tethered to RNA polymerase II promoters by a GAL4 DNA-binding domain actively represses transcription in a distance-independent manner. KRAB-mediated repression is dependent on the dose of the GAL4-KRAB-A fusion protein and on the presence of GAL4 binding sites on the DNA. Conversely, the LeR domain did not modulate significantly the transcription. Our results indicate that the KRAB domain present in the non-finger region of many ZFP genes quenches transcription possibly due to specific protein-protein interactions between the KRAB-A domain and components of the proximal transcriptional apparatus.
Assuntos
Sequência Conservada , Regulação da Expressão Gênica , Proteínas de Saccharomyces cerevisiae , Fatores de Transcrição , Transcrição Gênica , Dedos de Zinco/genética , Sequência de Aminoácidos , Sequência de Bases , Sítios de Ligação , Cloranfenicol O-Acetiltransferase/genética , DNA/metabolismo , Proteínas de Ligação a DNA , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Leucina , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão , Sequências Reguladoras de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico , TransfecçãoRESUMO
Starting from sequences of at least 60 bp, PCR-based screening has been developed to recover cDNAs from libraries without the necessity for hybridization or extensive DNA extraction steps. The method maintains the indefinite availability of even scarce cDNA libraries and provides an estimate of the relative abundance of the mRNA species. Isolation of a cDNA clone can be done in less than a week. cDNAs were isolated that were cognate for fragments of expressed sequences and for an exon predicted from genomic sequence.
Assuntos
Clonagem Molecular/métodos , DNA Complementar/genética , Reação em Cadeia da Polimerase/métodos , Animais , Cricetinae , Expressão Gênica , Biblioteca Gênica , HumanosRESUMO
The human genome contains a variety of genetic elements similar in structure to retroviruses and retrotransposons. We report here the structural and functional organization of a novel human endogenous retroviral family (ERV9). Three polyadenylated RNAs, 8, 2, and 1.5 kb long, are detected by Northern blot in undifferentiated embryonal carcinoma NT2/D1 cells. Upon genomic cloning of an expressed ERV9 locus, we demonstrated that the three polyadenylated RNAs are originated by a single ERV9 locus by alternative usage of splicing and polyadenylation signals. DNA sequence analysis of different ERV9 LTRs have revealed that they are heterogeneous in length and that the length variability is due to the number of tandemly repeated subelements present in both U3 and U5 regions; moreover, the ERV9 LTRs are capable to drive expression of a reporter gene in transient expression assays. Finally, analysis of the ERV9 5' transcription start site has allowed us to define the U3-R-U5 organization of the ERV9 LTR.
Assuntos
Retroviridae/genética , Sequência de Bases , Northern Blotting , Clonagem Molecular , DNA Viral , Humanos , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Células Tumorais CultivadasRESUMO
We have identified new repeated interspersed DNA sequences by analysis of homologous RNA transcripts from a human teratocarcinoma cell line (NTERA-2 clone D1). The abundance of transcripts varies upon retinoic acid induced differentiation of NTERA-2/D1 cells, and it is highest when the cells display the embryonal carcinoma phenotype. The expression of these novel repeated sequences appears to be tissue specific as no detectable expression was found in various cell lines of different embryological derivation. Characterization of the RNA transcripts by analysis of recombinant cDNA clones indicated that transcripts of different genomic units are present in undifferentiated embryonal teratocarcinoma cells. Nucleotide sequencing of the cloned cDNAs reveals a complex structure composed by unique and tandemly repeated sub-elements.
Assuntos
DNA/genética , Sequências Repetitivas de Ácido Nucleico , Teratoma/genética , Sequência de Bases , Códon , Regulação da Expressão Gênica , Humanos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , RNA/análise , RNA/genética , Mapeamento por Restrição , Transcrição Gênica , Tretinoína/farmacologia , Células Tumorais CultivadasRESUMO
We have isolated a novel full-length cDNA for a gene (IGSF1) located in distal Xq25. This transcript is highly expressed in adult testis and fetal liver but is undetectable in adult liver. A smaller alternate form is highly expressed in adult heart. The gene encodes a protein of 1327 amino acids with several recognizable functional domains. The protein has a putative signal peptide and transmembrane region, 15 potential sites for N-linked glycosylation, and 12 C2-type immunoglobulin (Ig)-like domains. All of the Ig-like domains contain the two conserved cysteine residues that form intradomain disulfide bonds typical of this superfamily. These features are consistent with a possible role for this molecule in cell surface recognition or cell-cell interaction.
Assuntos
Genes de Imunoglobulinas/genética , Imunoglobulinas/genética , Proteínas de Membrana/genética , Cromossomo X/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Biblioteca Gênica , Humanos , Imunoglobulinas/biossíntese , Imunoglobulinas/química , Proteínas de Membrana/biossíntese , Proteínas de Membrana/química , Modelos Moleculares , Dados de Sequência Molecular , Especificidade de Órgãos/genética , TeratocarcinomaRESUMO
We have isolated a full-length cDNA corresponding to the XAP5 gene in Xq28. An unusual feature of the cDNA is that it contains runs of CCG repeats in the 5' untranslated region, typical of genes that exhibit anticipation. It has a striking pattern of differential expression and is greatly enhanced in various fetal tissues. This predicted protein encodes a unique 339-amino-acid polypeptide that contains a large percentage of highly charged residues and a possible nuclear localization signal. A comparison to genomic sequence shows that XAP-5 comprises 13 exons spanning 6.5 kb. An examination of the human population indicates that the longest CCG run is polymorphic and varies in length from 8 to 12 repeats.
Assuntos
Proteínas Nucleares/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Complementar , Humanos , Dados de Sequência Molecular , Polimorfismo Genético , Repetições de Trinucleotídeos , Cromossomo XRESUMO
The evolutionarily conserved protein domain, called the Krüppel-associated box (KRAB), present in the amino termini of a large number of Krüppel-type zinc finger proteins is a strong repressor domain. In order to develop novel strategies to control human immunodeficiency virus type 1 (HIV-1) gene expression, we constructed a series of expression vectors expressing the wild-type Tat or Tat transdominant negative mutants fused to the KRAB repressor domain. We found that the KRAB domain tethered to the transactivating response element is able to suppress both basal and Tat-mediated activity of HIV-1 long terminal repeat-driven gene expression. These results suggest that the KRAB repressor domain fused to the Tat transdominant negative mutants can be successfully employed to control HIV-1 gene expression.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Regulação Viral da Expressão Gênica , Repetição Terminal Longa de HIV , HIV-1/genética , HIV-1/metabolismo , Regiões Promotoras Genéticas , Proteínas Repressoras/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica , Sequência de Aminoácidos , Evolução Biológica , Linhagem Celular , Cloranfenicol O-Acetiltransferase/biossíntese , Sequência Conservada , Produtos do Gene tat/biossíntese , Produtos do Gene tat/metabolismo , Células HeLa , Humanos , Fatores de Transcrição Kruppel-Like , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Transfecção , Células Tumorais Cultivadas , Dedos de Zinco , Produtos do Gene tat do Vírus da Imunodeficiência HumanaRESUMO
The human 3p21-22 region is frequently involved in karyotype rearrangements associated with malignancies. The high frequency of allelic loss in this region has been associated with virtually all small cell lung carcinomas and many renal carcinomas. These findings suggest that at least one tumor-suppressor gene might be located in 3p21-22. We have recently reported the isolation of a 750-kb yeast artificial chromosome (YAC) contig from 3p21-22. Here, we describe three new genes isolated from the 3p YAC contig by using a cDNA hybridization selection. Remarkably, the three new genes encode zinc-finger proteins, indicating the presence of a cluster of zinc-finger genes in human chromosome 3p21.
Assuntos
Cromossomos Humanos Par 3 , Família Multigênica , Dedos de Zinco/genética , Adulto , Mapeamento Cromossômico , Cromossomos Artificiais de Levedura , Clonagem Molecular , DNA Complementar , Expressão Gênica , Humanos , Hibridização in Situ Fluorescente , Transcrição GênicaRESUMO
Anhidrotic ectodermal dysplasia (EDA) is a disorder characterized by poor development of hair, teeth, and sweat glands, and results from lesions in the X-linked EDA gene. We have cloned a 1.6-kilobase 5'-flanking region of the human EDA gene and used it to analyze features of transcriptional regulation. Primer extension analysis located a single transcription initiation site 264 base pairs (bp) upstream of the translation start site. When the intact cloned fragment or truncated derivatives were placed upstream of a reporter luciferase gene and transfected into a series of cultured cells, expression comparable with that conferred by an SV40 promoter-enhancer was observed. The region lacks a TATA box sequence, and basal transcription from the unique start site is dependent on two binding sites for the Sp1 transcription factor. One site lies 38 bp 5' to the transcription start site, in a 71-bp sequence that is sufficient to support up to 35% of maximal transcription. The functional importance of the Sp1 sites was demonstrated when cotransfection of an Sp1 expression vector transactivated the EDA promoter in the SL2 Drosophila cell line that otherwise lacks endogenous Sp1. Also, both Sp1 binding sites were active in footprinting and gel shift assays in the presence of either crude HeLa cell nuclear extract or purified Sp1 and lost activity when the binding sites were mutated. A second region involved in positive control was localized to a 40-bp sequence between -673 and -633 bp. This region activated an SV40 minimal promoter 4- to 5-fold in an orientation-independent manner and is thus inferred to contain an enhancer region.
Assuntos
Displasia Ectodérmica/genética , Ligação Genética , Regiões Promotoras Genéticas , Cromossomo X , Sequência de Bases , Linhagem Celular , DNA , Pegada de DNA , Ectodisplasinas , Elementos Facilitadores Genéticos , Humanos , Proteínas de Membrana/genética , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Sequências Reguladoras de Ácido Nucleico , Fator de Transcrição Sp1/metabolismoRESUMO
A novel endogenous retroviral sequence (ERV-9) has been isolated from a human embryonal carcinoma cDNA library by hybridization to a probe containing a recently described human repetitive element. DNA sequence analysis of the 4kb cDNA insert (pHE.1) revealed the presence of ORFs potentially coding for putative retrovirus-related gag, pol and env proteins. Northern blot and RNase protection experiments showed that RNA homologous to the pHE.1 insert is detected only in embryonal carcinoma cells as a 8 kb mRNA, and its expression is negatively regulated during retinoic acid induced differentiation of the human teratocarcinoma cell line NT2/D1. Using a pol specific probe we have isolated a genomic locus containing the ERV-9 sequences. Characterization by restriction enzyme analysis and DNA sequencing allowed us to define LTR-like sequences, that are composed by a complex array of subrepetitive elements. In addition we show that ERV-9 LTR sequences are capable to drive expression of linked CAT gene in a cell specific manner as LTR promoter activity has been detected only in NT2/D1 cells.
Assuntos
Células-Tronco Neoplásicas/metabolismo , Retroviridae/genética , Sequência de Aminoácidos , Sequência de Bases , Southern Blotting , DNA de Neoplasias/análise , DNA de Neoplasias/genética , DNA Viral/análise , DNA Viral/genética , Células-Tronco de Carcinoma Embrionário , Genes Virais , Humanos , Dados de Sequência Molecular , Fases de Leitura Aberta , Plasmídeos , Regiões Promotoras Genéticas , RNA Neoplásico/análise , RNA Neoplásico/genética , RNA Viral/análise , RNA Viral/genética , Mapeamento por Restrição , Alinhamento de Sequência , Células Tumorais CultivadasRESUMO
It has been demonstrated previously that the transcriptional repressor domain called the Krüppel-associated box (KRAB), conserved in a large number of Krüppel-type zinc finger proteins, fused to Tat transdominant negative mutants, is able to silence HIV-1 long terminal repeat (LTR)-driven gene expression in transient transfection assays. In the present study chimeric Tat mutant-KRAB retroviral expression vectors were used to control HIV-1 replication in acutely infected cells. It was found that while transient and stable expression of Tat mutant-KRAB chimeric proteins represses HIV-1 LTR-driven gene transcription in transient assays, stable expression of Tat mutant-KRAB chimeric molecules does not confer resistance to HIV-1 infection in Jurkat T lymphocytic cell lines. The results provide further evidence that transient transfection may underestimate the role of chromosomal structure in transcriptional regulation and highlight the caveat of direct extrapolation of transient results for designing gene therapy strategies for efficient control of HIV-1 infection.
Assuntos
Proteínas de Ligação a DNA/fisiologia , Repetição Terminal Longa de HIV/genética , Proteínas Repressoras/fisiologia , Fatores de Transcrição/fisiologia , Animais , Linhagem Celular , Proteínas de Ligação a DNA/genética , Regulação Viral da Expressão Gênica , Produtos do Gene tat/genética , Células HeLa , Humanos , Células Jurkat , Fatores de Transcrição Kruppel-Like , Mutação , Proteínas Recombinantes de Fusão/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Transcrição Gênica , Transfecção , Integração Viral , Dedos de ZincoRESUMO
The region surrounding the ZNF35 zinc finger protein gene on 3p21 is of particular interest, as this region of chromosome 3 is frequently involved in rearrangements and/or deletions associated with various human tumors including lung and renal carcinoma. We have analyzed yeast artificial chromosomes (YACs), identified by PCR screening, using oligonucleotides derived from the ZNF35 gene. PFGE and Southern blot/hybridization analysis revealed that the clones cover 750-kb including the ZNF35 gene. The use of specific somatic cell hybrids have allowed us to locate the YAC contig telomeric to the D3F15S2 locus, in a region which is frequently deleted in lung carcinomas. In addition, we have developed a novel cDNA hybridization protocol allowing the isolation of transcribed sequences present in the overlapping YAC clones. Using the cDNA hybridization selection, we have isolated and characterized one transcribed sequence (D3S1362E) from the 3p21 YAC contig and the corresponding cDNA has been isolated. DNA sequencing analysis indicated that the D3S1363E cDNA codes for a putative transcription factor. Northern blot analysis indicated that the D3S1362E sequence hybridized to multiple transcripts in skeletal muscle, and weakly hybridizing transcripts of similar sizes were detected in other tissues.
Assuntos
Cromossomos Fúngicos , Cromossomos Humanos Par 3/ultraestrutura , Biblioteca Gênica , Genoma Humano , Transcrição Gênica , Dedos de Zinco/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Passeio de Cromossomo , Clonagem Molecular , Sequência Consenso , Genes Supressores de Tumor , Humanos , Células Híbridas , Camundongos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
On the basis of sequence similarity in the repeated zinc finger domain, we have identified and characterized two human cDNA clones (ZNF7 and ZNF8), both encoding proteins containing potential finger-like nucleic acid binding motifs. Northern blot analysis indicates that both genes are expressed as multiple transcripts and they are ubiquitously present in many human cell lines of different embryological derivation. Moreover, their expression is modulated during in vitro induced terminal differentiation of human myeloid cell line HL-60. By in situ hybridization experiments, we have localized the ZNF7 gene to chromosome 8 (region q24) and the ZNF8 gene to the terminal band of the long arm of chromosome 20 (20q13).
Assuntos
Cromossomos Humanos Par 20 , Cromossomos Humanos Par 8 , Proteínas de Ligação a DNA/genética , Expressão Gênica , Genes , Metaloproteínas/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Diferenciação Celular , Linhagem Celular , Mapeamento Cromossômico , Clonagem Molecular , DNA/genética , DNA/isolamento & purificação , Feminino , Humanos , Fatores de Transcrição Kruppel-Like , Dados de Sequência Molecular , Homologia de Sequência do Ácido Nucleico , Transcrição Gênica , ZincoRESUMO
We report the structural and functional characterization of the HF.10 zinc finger gene (ZNF35) in normal human cells, as well as a processed pseudogene. The HF.10 gene spans about 13 kb and it is interrupted by three introns. All 11 zinc finger DNA-binding domains are contiguously encoded within the last 3' exon. The genomic region surrounding HF.10 exon 1 contains a CpG island and acts as a promoter in vitro. Using transient CAT assay in cotransfection experiments in cultured cells, we have determined that the HF.10 finger protein is a transcriptional transactivator. Restriction enzyme mapping and partial nucleotide sequencing of the HF.10 pseudogene indicated that it has arisen by retroposition of spliced HF.10 mRNA. In situ hybridization experiments revealed that both the functional locus and the pseudogene map to chromosome 3p21p22, a region that is frequently deleted in small cell lung and renal carcinomas. Hybridization of the HF.10 gene and the HF.10 pseudogene DNA probes to metaphases from a small cell lung carcinoma cell line with the 3p deletion revealed that both loci are part of the deleted chromosome region.
Assuntos
Cromossomos Humanos Par 3 , Dedos de Zinco/genética , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , DNA/genética , Genes , Humanos , Dados de Sequência Molecular , Pseudogenes , Transativadores/genética , Transcrição GênicaRESUMO
The finger motif is a tandemly repeated DNA-binding domain recently identified in the primary structure of several eukaryotic transcriptional regulatory proteins. It has been proposed that some members of the finger-gene family are implicated in both normal cell proliferation and differentiation. We isolated several human finger genes by means of hybridization with a finger motif-containing DNA probe. One of these finger genes, HF.10, is expressed at low levels in a variety of human tissues and is down-regulated during the in vitro terminal differentiation of human leukemic myeloid cell lines. By in situ hybridization experiments and analysis of interspecific somatic cell hybrids we mapped the HF.10 gene to 3p21-22, a chromosome region frequently involved in karyotypic rearrangements associated with lung and renal cancer.
Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 3 , Proteínas de Ligação a DNA/genética , Metaloproteínas/genética , Neoplasias/genética , Animais , Southern Blotting , Mapeamento Cromossômico , Sondas de DNA , Humanos , Células Híbridas , Camundongos , Família MultigênicaRESUMO
The Oct-2 transcription factor contains an inhibitory domain which is able to repress transcription following DNA binding. Here we show that within the neuronally expressed Oct-2.5 form, the inhibitory domain can strongly inhibit activation by transcription factor activation domains which are either composed predominantly of acidic residues or contain the HOB motif, whereas it has a weaker effect or no effect on proline-rich activation domains and on a glutamine-rich domain. In contrast, the isolated inhibitory domain of Oct-2 can efficiently repress all types of activation domains. This effect is observed however, only on TATA box-containing promoters and not on promoters containing an initiator motif. This widespread inhibition of different activation domains and its dependence on the nature of the basal promoter elements indicate that the inhibitory domain is likely to act by contacting a common downstream target of activation domains within the basal transcriptional complex bound at the TATA box rather than quenching specific activation domains by direct interaction. These effects are discussed in terms of the functional role of the inhibitory domain within Oct-2.5 and the mechanism by which it acts.
Assuntos
Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica , Regiões Promotoras Genéticas , Proteínas Repressoras/genética , Proteínas de Saccharomyces cerevisiae , Fatores de Transcrição , Transcrição Gênica , Ativação Transcricional , Animais , Células Cultivadas , Cricetinae , Proteínas Fúngicas/genética , RNA Mensageiro/genética , Proteínas Recombinantes , TransfecçãoRESUMO
cDNA clones encoding zinc finger motifs were isolated by screening human placenta and T-cell (Peer) cDNA libraries with zinc finger (ZNF) consensus sequences. Unique cDNA clones were mapped in the human genome by rodent-human somatic cell hybrid analysis and in some cases in situ chromosomal hybridization. ZNF80 mapped to 3p12-3qter, ZNF7 was previously mapped to 8q24 and is here shown by in situ hybridization and use of appropriate hybrids to map telomeric to the MYC locus. ZNF79 mapped to 9q34 centromeric to the ABL gene and between a constitutional chromosomal translocation on the centromeric side and the CML specific ABL translocation on the telomeric side. ZNF77 mapped to 19p while ZNF78L1 (pT3) mapped to 19q. Chromosome 19 carries many ZNF loci and other genes with zinc finger encoding motifs; the pT3 clone additionally detected a locus designated ZNF78L2, which mapped to chromosome region 1p, most likely in the region 1p32 where the MYCL and JUN loci map.
Assuntos
Mapeamento Cromossômico , Dedos de Zinco , Sequência de Aminoácidos , Animais , Cromossomos Humanos Par 19 , Cromossomos Humanos Par 3 , Cromossomos Humanos Par 8 , Cromossomos Humanos Par 9 , Clonagem Molecular , Humanos , Células Híbridas , Dados de Sequência Molecular , Família MultigênicaRESUMO
A yeast artificial chromosome sequence-tagged site-based (YAC/STS) physical map of 22.5 Mb of the Xq24-q26 cytogenetic band region of the human X chromosome has been assembled. DNA coverage includes 857 large-insert clones formatted with 405 STSs to provide ninefold depth of DNA. At five points, no bridging clones have been recovered from 20 X-chromosome equivalents of human DNA in YACs or bacterial clones, but the placement of 25 ("CA")n polymorphic markers permits the ordering of contigs by comparison with the genetic linkage map and radiation hybrid data. The map localizes the X3000 translocation breakpoint and six genes (ANT2, NDUFA1, LAMP2, OCRL, IGSF1, and HDGF) at better than 100-kb resolution. The relatively gene-poor nature of the region is consistent with relatively low uniform 34-42% GC content in STSs across nearly all of the region.