RESUMEN

Homeodomain transcription factors play important roles in directing cellular proliferation and differentiation. A TALE-superclass homeodomain protein, multifunctional repressor of TGFbeta-induced transcription. Here we report identification of TGIF2, a novel TALE-superclass homeodomain protein that shows distinct homology with TGIF, especially in its DNA-binding domain. TGIF2 is expressed ubiquitously in human tissues, with the highest levels being found in heart, kidney, and testis. The TGIF2 product contains a putative nuclear localization signal; translocation of the protein to the nucleus was confirmed by transfection of epitope-tagged cDNA. TGIF2 lies on chromosome 20q11.2-12. Since amplification of 20q is often observed among ovarian cancers, we determined the status of DNA copy-number and expression of TGIF2 in 14 ovarian-cancer cell lines. This gene was over-expressed in all lines that showed amplification by FISH analysis. The results suggested that TGIF2 may play an important role in the development and/or progression of some ovarian tumors through a mechanism of gene amplification.

Asunto(s)

Amplificación de Genes , Regulación Neoplásica de la Expresión Génica , Genes Homeobox , Proteínas de Homeodominio/genética , Neoplasias Ováricas/genética , Secuencia de Aminoácidos , Femenino , Humanos , Datos de Secuencia Molecular , Proteínas Represoras/genética , Alineación de Secuencia , Células Tumorales Cultivadas

RESUMEN

Members of the T-box family are known to play critical roles in the embryonic development of most animal species. Recently, we have isolated its new mammalian member, Tbr2, from mouse embryonic brain. We have also shown that the expression patterns of Tbr2 and the closely related Tbr1 appear to be reciprocal in the developing brain; Tbr2 is expressed in mesencephalon and rhombencephalon, but expression of Tbr1 is restricted to telencephalon. To investigate possible structural and functional relationships of Tbr2 and other T-box containing genes, we analyzed genomic organization of the murine Tbr2 gene. The Tbr2 gene is composed of six exons (1353, 155, 122, 159, 62 and 1035bp), and five introns (920, 643, 602, 85 and 2036bp). This exon/intron organization is very similar to that of Tbr1. We also analyzed the 3.9kb sequence of the 5' promoter region flanking the Tbr2 gene and the corresponding region of the Tbr1 gene. The sites for Brn-2 and Tst-1 were found in the promoter of Tbr2 but not Tbr1. On the contrary, there were eight HNF-3beta binding sites in the Tbr1 gene promoter but only three in the Tbr2 promoter. The differential presence of putative binding sites for these brain-specific transcription factors may explain the reciprocal expression of Tbr1 and Tbr2. Furthermore, a single chromosomal locus for mouse Tbr2 was assigned to 9F3 by fluorescence in-situ hybridization 1.

Asunto(s)

Proteínas de Unión al ADN/genética , Genes/genética , Proteínas del Tejido Nervioso , Proteínas de Dominio T Box , Animales , Secuencia de Bases , Sitios de Unión , Mapeo Cromosómico , ADN/química , ADN/genética , Exones , Hibridación Fluorescente in Situ , Intrones , Ratones , Ratones Endogámicos , Datos de Secuencia Molecular , Regiones Promotoras Genéticas/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , TATA Box

RESUMEN

Chromosome 4p- syndrome is a multiple malformation syndrome associated with partial deletion of the short arm of chromosome 4 (4p-). It is characterized by dysmorphic features and retarded development. Cleft lip and/or palate are the major clinical manifestations. Cases of tetrasomy 9p are extremely rare; the principal clinical manifestations of this condition are characteristic craniofacial abnormalities, generalized hypotonia and severe mental retardation. We present the first case of a female infant with 4p deletion and tetrasomy 9p mosaicism, exhibiting a left-sided cleft lip, alveolus and soft palate. Karyotype analysis of lymphocytes cultured from the patient revealed that she was mosaic: 86% of the cells were 46, XX, add (4) (p15.32) and 14% were 47, XX, add (4) (p15.32), +idic (9)(q12). The G-banding pattern appeared consistent with either translocation or partial proximal deletion of 4p. In order to make a definitive cytogenetic diagnosis of isodicentric chromosome 9, fluorescence in situ hybridization (FISH) was applied. At 8 months, when the patient weighed 4.3 kg, her cleft lip was repaired. Before and after surgery there were no seizures, and the postoperative course was uneventful.

Asunto(s)

Anomalías Múltiples/genética , Aberraciones Cromosómicas/genética , Cromosomas Humanos Par 4 , Cromosomas Humanos Par 9 , Labio Leporino/genética , Fisura del Paladar/genética , Deleción Cromosómica , Trastornos de los Cromosomas , Femenino , Humanos , Lactante , Discapacidad Intelectual/genética , Isocromosomas , Cariotipificación , Poliploidía , Síndrome , Translocación Genética

RESUMEN

Caspase-activated DNase (CAD) cleaves chromosomal DNA during apoptosis. We determined its genomic structure and identified single-nucleotide polymorphisms (SNPs) within exons 5 and 7, as well as a highly polymorphic dinucleotide repeat of (CT)m(CA)n within the 5' region of the human CAD gene (hCAD). The genomic structure of hCAD presented here, together with information concerning SNPs within the gene, as well as a highly polymorphic (CT)m(CA)n repeat fragment at the hCAD locus, may assist in the construction of genetic maps for exploring gene(s) that play pivotal roles in carcinogenesis.

Asunto(s)

Desoxirribonucleasas/genética , Repeticiones de Dinucleótido/genética , Polimorfismo de Nucleótido Simple , Exones/genética , Humanos

RESUMEN

KNSL4 (Kid; kinesin-like DNA-binding protein) is a member of the kinesin family that is involved in spindle formation and the movements of chromosomes during mitosis and meiosis. Myc-associated zinc finger protein (MAZ) participates in both the initiation and the termination of transcription of target genes. We isolated genomic DNA clones that encoded KNSL4 and MAZ from a human cosmid library. Sequence analysis revealed that the two genes were very close to one another. The distance between the two genes was only 1. 2 kb, and this intervening 1.2-kb region was extremely GC-rich. The gene for KNSL4 spanned 16 kb and consisted of 14 exons and 13 introns, while the gene for MAZ spanned 6 kb and consisted of 5 exons and 4 introns. The two genes were mapped to chromosome 16p11.2 by fluorescence in situ hybridization.

Asunto(s)

Cromosomas Humanos Par 16/genética , Proteínas de Unión al ADN/genética , Cinesinas/genética , Factores de Transcripción/genética , Mapeo Cromosómico , Clonación Molecular , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Empalme del ARN/genética , Mapeo Restrictivo , Análisis de Secuencia de ADN , Dedos de Zinc/genética

Asunto(s)

Mapeo Cromosómico/métodos , Cromosomas Humanos Par 10 , Cromosomas Humanos Par 4 , Proteínas Serina-Treonina Quinasas/genética , Receptores de Factores de Crecimiento/genética , Animales , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1 , Cartilla de ADN , Marcadores Genéticos , Humanos , Hibridación Fluorescente in Situ , Ratones , Reacción en Cadena de la Polimerasa

RESUMEN

The cDNA for the seventh mammalian aquaporin (AQP7) was isolated from rat testis, and its expression demonstrated at the tail of late spermatids (Ishibashi et al., J. Biol. Chem. 272 (1997) 20,782-20,786). Here we report the isolation of the mouse and the human AQP7 cDNA and the human AQP7 gene. The human AQP7 gene is identical with human adipose AQP (AQPap or AQP7L). The deduced amino acid sequences of human and mouse AQP7 were 68% and 79% identical to those of rat AQP7, respectively. The mouse AQP7 is 67% identical to the human AQP7. Such a lower conservation of AQP7 among species is unusual in the aquaporin family. The human AQP7 gene is composed of six exons distributing over 6.5 kb. The exon-intron boundaries are identical to those of the human AQP3 gene. The intron sizes are also similar. Moreover, chromosomal localization of AQP7 was assigned to 9p13 by fluorescent in situ hybridization, where AQP3 is also localized, suggesting that 9p13 may be another site of an aquaporin cluster.

Asunto(s)

Acuaporinas , Canales Iónicos/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Mapeo Cromosómico , Clonación Molecular , ADN Complementario/química , ADN Complementario/aislamiento & purificación , Humanos , Canales Iónicos/química , Masculino , Ratones , Datos de Secuencia Molecular , Alineación de Secuencia , Testículo/metabolismo

RESUMEN

Members of the ATP binding cassette (ABC) superfamily are involved in the energy-dependent transport of a wide variety of substrates including anticancer agents across the membranes. We have cloned a cDNA fragment including a novel ABC sequence from a cisplatin-resistant human lung adenocarcinoma cell line, PC-14/CDDP, by reverse transcription polymerase chain reaction (RT-PCR) using degenerate primers and screened a cDNA library using the cDNA fragment as a probe. A full-length cDNA clone, BM4.8, was obtained. Sequence analysis showed that the cDNA encoded a short type of multidrug resistance protein homologue, SMRP, by computed homology search. SMRP was composed of 946 amino acids and had two ABCs with walker A and B motifs. This gene was mapped on chromosome 3 at band q27 by fluorescence in situ hybridization (FISH) analysis and was found to be expressed in various tissues by Northern blot analysis.

Asunto(s)

Transportadoras de Casetes de Unión a ATP/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , ADN Complementario/aislamiento & purificación , Resistencia a Múltiples Medicamentos , Neoplasias Pulmonares/genética , Proteínas de Neoplasias/genética , Transportadoras de Casetes de Unión a ATP/aislamiento & purificación , Secuencia de Aminoácidos , Secuencia de Bases , Carcinoma de Pulmón de Células no Pequeñas/química , Mapeo Cromosómico , Cromosomas Humanos Par 3 , Cisplatino/farmacología , Clonación Molecular , Humanos , Neoplasias Pulmonares/química , Datos de Secuencia Molecular , Proteínas Asociadas a Resistencia a Múltiples Medicamentos , Familia de Multigenes , Proteínas de Neoplasias/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Células Tumorales Cultivadas

Asunto(s)

Acil-CoA Deshidrogenasas/genética , Mapeo Cromosómico , Acil-CoA Deshidrogenasa de Cadena Larga , Acil-CoA Deshidrogenasas/metabolismo , Animales , Sitios de Unión , Clonación Molecular , Cartilla de ADN , Biblioteca de Genes , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Ratas , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADN , TATA Box , Transcripción Genética

RESUMEN

The transcription factor KBF2 has been characterized as a factor that binds to the NFkB site of mouse major histocompatibility complex (MHC) class I genes and its amino acid sequence has been shwn to be identical to those of members of the recombination signal-sequence binding protein (RBP-Jk) family. Previous studies by Amakawa et al. (Genomics 17, 306-315, 1993) demonstrated that the functional gene is localized at human chromosome 3q25. However, in the present study we showed by in situ hybridization with the functional KBF2/RBPJk cosmid clone that the gene is localized at 9p12-13 and 9q13, namely, at the same loci as pseudogenes that were reported previously (Zhang et al, Jpn J Human Genet 39, 391-401, 1994).

Asunto(s)

Cromosomas Humanos Par 9 , Proteínas de Unión al ADN/genética , Proteínas Nucleares , Animales , Cósmidos , Humanos , Proteína de Unión a la Señal Recombinante J de las Inmunoglobulinas , Hibridación Fluorescente in Situ , Ratones , Factores de Transcripción/genética

Asunto(s)

Hemo Oxigenasa (Desciclizante)/genética , Isoenzimas/genética , Ratones/genética , Animales , Bandeo Cromosómico , Mapeo Cromosómico , Sondas de ADN , Hemo-Oxigenasa 1 , Hibridación Fluorescente in Situ , Proteínas de la Membrana

RESUMEN

Members of the nuclear factor of activated T cells (NFAT) are involved in the induction of a number of cytokine genes. We report here cDNA cloning and chromosomal localization of a murine homologue of human NFATx, designated as mNFATx1, and its splicing variants mNFATx2 and m delta NFATx. Northern blot analysis showed mNFATx1 to be predominantly expressed in the thymus. mNFATx1, but not m delta NFATx, produced in COS-7 cells, bound to all NFAT-binding sites of the interleukin (IL)-2 and IL-4 promoters tested. Immunofluorescence assay showed that both mNFATx1 and m delta NFATx introduced into COS-7 cells localized predominantly to the cytoplasm, but did translocate to the nucleus, either by cotransfection with an active form of calcineurin or wild-type calcineurin followed by stimulation with calcium ionophore. Translocation of mNFATx1 correlated well with activation of the murine IL-2 promoter; mNFATx1 translocated under conditions described above, in combination with phorbol 12-myristate 13-acetate, activated the transiently transfected murine IL-2 promoter. Thus, nuclear-translocated mNFATx1 is involved in activation of the IL-2 promoter. These results provide the first evidence for the requirement of calcineurin in the control of mNFATx imported from the cytoplasm to the nucleus and implies that mNFATx may possibly be a substrate of calcineurin in vivo.

Asunto(s)

Proteínas de Unión a Calmodulina/metabolismo , Núcleo Celular/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteínas Nucleares , Fosfoproteínas Fosfatasas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Sitios de Unión , Transporte Biológico , Northern Blotting , Células COS/metabolismo , Calcineurina , Mapeo Cromosómico , Cromosomas , Clonación Molecular , Femenino , Humanos , Interleucina-2/genética , Interleucina-2/metabolismo , Interleucina-4/genética , Interleucina-4/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Factores de Transcripción NFATC , Regiones Promotoras Genéticas , Empalme del ARN , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Transducción de Señal , Distribución Tisular , Transcripción Genética , Transfección

Asunto(s)

Acil-CoA Deshidrogenasas/genética , Secuencia Conservada , Acil-CoA Deshidrogenasa de Cadena Larga , Animales , Secuencia de Bases , Bandeo Cromosómico , Mapeo Cromosómico , Femenino , Humanos , Hibridación Fluorescente in Situ , Ratones , Ratones Endogámicos ICR , Ratones Desnudos , Especificidad de la Especie

RESUMEN

A mouse cDNA encoding a human homologue of heme oxygenase-2 (HO-2) was isolated. The deduced protein contains 315 amino acids and has a calculated molecular mass of 35.8 kDa. The nucleotide sequence is 85.6% identical and the amino acid sequence 87% identical to those of the human protein. The corresponding mRNA is present in brain and testis, but not in ovary, kidney, liver, or spleen.

Asunto(s)

ADN Complementario/química , Hemo Oxigenasa (Desciclizante)/genética , Isoenzimas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Humanos , Ratones , Datos de Secuencia Molecular , Peso Molecular , Alineación de Secuencia

Asunto(s)

Canales de Cloruro/genética , Cromosomas Humanos Par 1 , Cromosomas Humanos Par 4 , Mapeo Cromosómico , Humanos , Hibridación Fluorescente in Situ

RESUMEN

The Down syndrome (DS) region on chromosome 21, which is responsible for the main features of DS such as characteristic facial features, a congenital heart defect, and mental retardation, has been defined by molecular analysis of DS patients with partial trisomy 21. The 2. 5-Mb region around the marker D21S55 between D21S17 and ERG in 21q22 is thought to be important, although contributions of other regions cannot be excluded. In this region, we focused on a 1.6-Mb region between a NotI site, LA68 (D21S396, which is mapped distal to D21S17) and ERG, because analysis of a Japanese DS family with partial trisomy 21 revealed that the proximal border of its triplicated region was distal to LA68. We constructed P1 contigs with 46 P1 clones covering more than 95% of the 1.6-Mb region. A high-resolution restriction map using BamHI was also constructed for more detailed analysis. Our P1 contig map supplements other physical maps previously reported and provides useful materials for further analysis including gene isolation and sequencing of the DS region.

Asunto(s)

Mapeo Cromosómico/métodos , Cromosomas Humanos Par 21 , Síndrome de Down/genética , Secuencia de Bases , Cromosomas Artificiales de Levadura , Cromosomas Humanos Par 4 , Cartilla de ADN/química , Marcadores Genéticos , Biblioteca Genómica , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Mapeo Restrictivo , Lugares Marcados de Secuencia , Translocación Genética

RESUMEN

A reliable method for obtaining high-resolution R-banded chromosomes from lymphoblastoid cell lines is described. The cell cultures are subjected to S-phase synchronization in the presence of excess thymidine (300 micrograms/ml) for 17 to 19 hr, followed by BrdU treatment (30 micrograms/ml) for 6.5 hr. Prior to harvest, they are exposed to ethidium bromide (7.5 micrograms/ml) for 1.5 hr and Colcemid (0.02 microgram/ml) for 30 min. Using this method, high-resolution R-banded chromosomes at the 550-850 band level were obtained with frequencies as high as 70% of all mitotic cells.

Asunto(s)

Bandeo Cromosómico/métodos , Mapeo Cromosómico , Translocación Genética , Ciclo Celular , Línea Celular , Etidio , Humanos , Linfocitos , Metotrexato , Células Tumorales Cultivadas