RESUMEN
An inv(11)(q13.5;q25) inversion was previously identified in a 9-month-old male patient with complex cyanotic heart defects, altered lung lobation, symmetric liver, and abnormally lobulated spleen (polysplenia). This chromosomal rearrangement was inherited from the phenotypically normal father. We termed these regions DHTX-A (disrupted in heterotaxy)-- A at 11q13.5 and DHTX-B at 11q25. Here, we report the isolation and characterization of the inversion breakpoints and the gene that is disrupted by the DHTX-A breakpoint. The putative DHTX is identical to the UVRAG gene, which was originally identified as a gene that complements the UV sensitivity of xeroderma pigmentosum complementation group C. The 4-kb mRNA was found to be encoded by a large gene, at least 300 kb long, composed of 15 exons. The function of the gene product remains largely unknown. However, the near central portion of the UVRAG protein is predicted to contain a coiled-coil domain, which has been implicated in mediating protein-protein interactions. Southern analyses and fluorescence in situ hybridization (FISH) revealed that the DHTX-A breakpoint in the patient and his father lies within the intron between exons 6 and 7 of UVRAG. Northern blot analysis indicated strong expression in human fetal and adult tissues and in mouse embryonic day-7 and adult tissues, respectively. Whole mount in situ hybridization also showed that the Uvrag gene is expressed in the presomite-stage embryo. Several hypotheses are discussed to explain the relationship between the chromosomal inversion and the accompanying phenotypes.
Asunto(s)
Aberraciones Cromosómicas/genética , Cromosomas Humanos Par 11 , Cardiopatías Congénitas/genética , Proteínas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Southern Blotting , Línea Celular , Inversión Cromosómica , Clonación Molecular , Dextrocardia/genética , Desarrollo Embrionario y Fetal , Exones , Cardiopatías Congénitas/diagnóstico por imagen , Humanos , Hibridación in Situ , Lactante , Intrones , Masculino , Ratones , Datos de Secuencia Molecular , Mapeo Físico de Cromosoma , Proteínas/química , ARN Mensajero/genética , Radiografía , Proteínas Supresoras de TumorRESUMEN
Shwachman syndrome is an autosomal-recessive disorder characterized by exocrine pancreatic insufficiency, bone-marrow dysfunction, and metaphyseal chondrodysplasia. A de novo balanced translocation was recently documented in a patient with this disease. Toward isolating the gene(s) responsible for Shwachman syndrome, we cloned and sequenced the translocation breakpoints in the DNA of this patient. The nucleotide sequences around the breakpoints contained neither repetitive elements nor motifs reported to be implicated in recombination events, although we did detect gains or losses of oligonucleotides at the translocation junctions. By large-scale genomic sequencing and in silico gene trapping, we identified two novel transcripts in the vicinity of the breakpoints that might represent candidate genes for Shwachman syndrome, one on chromosome 6 and the other on chromosome 12. The gene on chromosome 12 was actually disrupted by the translocation.
Asunto(s)
Insuficiencia Pancreática Exocrina/genética , Translocación Genética , Secuencia de Aminoácidos , Secuencia de Bases , Cromosomas Artificiales de Levadura , Cromosomas Humanos Par 12 , Cromosomas Humanos Par 6 , Clonación Molecular , ADN Complementario/análisis , Humanos , Cariotipificación , Datos de Secuencia Molecular , Insuficiencia Multiorgánica/genética , Sistemas de Lectura Abierta , Homología de Secuencia de Ácido Nucleico , SíndromeRESUMEN
From a human pancreas cDNA library we isolated and characterized a novel zinc finger gene encoding a protein homologous to ZNF187, a serum response element-binding protein. The full-length cDNA contained an open reading frame of 1,686 nucleotides encoding a predicted 562-amino-acid peptide that included an ATP-GTP binding site and seven C2H2 zinc finger domains. The consensus sequence of the C2H2 domains (CX2CX3FX5LX2HX3H) is common in the SRE-binding region present in Drosophila Krüppel proteins. An alternatively spliced form of the transcript found in the cDNA library lacked both the ATP-GTP binding site and any C2H2 zinc finger domains. We localized this gene (ZNF188) to chromosome band 7q22.1-->q22.3 by fluorescence in situ hybridization.
Asunto(s)
Mapeo Cromosómico , Proteínas de Unión al ADN/genética , Proteínas Represoras , Dedos de Zinc/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromosomas Humanos Par 7/genética , Clonación Molecular , ADN Complementario/genética , Proteínas de Unión al ADN/metabolismo , Drosophila/genética , Proteínas de Drosophila , Humanos , Hibridación Fluorescente in Situ , Proteínas de Insectos/genética , Factores de Transcripción de Tipo Kruppel , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Homología de Secuencia de Aminoácido , Factor de Respuesta Sérica , Factores de Transcripción/genéticaRESUMEN
Organogels of low molecular weight compounds, N-benzyloxycarbonyl-l-alanine 4-hexadecanoyl-2-nitrophenyl ester (BLAHN) and related compounds, in methanol and cyclohexane solvents were studied by FT-IR, circular dichroism (CD), transmission electron microscope (TEM), and scanning electron microscope (SEM). The gels exhibited a thermally reversible sol-to-gel phase transition, which gave the thermodynamic parameters for the transition. The FT-IR spectroscopic results suggested that the intermolecular hydrogen bonding between N-H and C=O of urethane bond played a vital role in gelation. The aggregates for gelation were mainly assembled by pi-pi interaction, dipole-dipole interaction, and the hydrophobic interaction as well as hydrogen bonding for BLAHN. The comparison of CD spectra of BLAHN, N-benzyloxycarbonyl-d-alanine 4-hexadecanoyl-2-nitrophenyl ester (BDAHN), and related molecules indicated that the self-aggregation of gelator molecules gradually proceeds and the component molecules are cooperatively organized to form a chiral aggregate. The three-dimensional interlocking structures and tree-branched structures responsible for the immobilization of fluid were observed by TEM and SEM. Copyright 1997 Academic Press. Copyright 1997Academic Press
RESUMEN
The human S1-5 gene (fibrillin-like; FBNL) was originally isolated from a subtractively enriched cDNA library established from a subject with Werner syndrome (WS). We isolated genomic clones containing the entire S1-5 gene and determined its genomic structure including the exon-intron organization. The gene spanned approximately 18 kb of genomic DNA and consisted of 12 exons. Its expression was abundant in all tissues examined except brain and peripheral leukocytes, where it was undetectable. In addition, we have mapped S1-5 by fluorescence in situ hybridization to chromosome 2p16, a position that excludes it as a candidate for WS. Our data should facilitate an understanding of the function and regulation of S1-5 in human tissues.
Asunto(s)
Cromosomas Humanos Par 2 , Proteínas de Microfilamentos/genética , Secuencia de Bases , Células Cultivadas , Mapeo Cromosómico , ADN Complementario , Exones , Fibrilinas , Expresión Génica , Humanos , Hibridación Fluorescente in Situ , Intrones , Datos de Secuencia Molecular , Distribución Tisular , Síndrome de Werner/genéticaRESUMEN
We were able to improve the success rate of blind nasotracheal intubation by using nasogastric tubes as a guide during intubation, first, for passing the endotracheal tube through the nasal cavity, and second, passing it from the pharynx to the larynx. By adding both sedation by modified neuroleptanalgesia (NLA) and topical and transtracheal administration of lidocaine, our technique became safer and smoother. We have completed 36 cases without accident, with an average time for intubation of 8.25 min. The Rüsh spiral tube was thought to be the most suited to this form of intubation because of the 90 degrees cut of its tip, its high-volume cuff, and its flexibility in all directions. These features are useful for hearing breath sounds, raising the tip of the tube by inflation of the cuff, and advancing the tube in a turning motion.
Asunto(s)
Anestesia Dental/instrumentación , Intubación Intratraqueal/instrumentación , Intubación Intratraqueal/métodos , Anestésicos Locales/administración & dosificación , Humanos , Intubación Gastrointestinal/instrumentación , Lidocaína/administración & dosificación , Neuroleptanalgesia/instrumentaciónRESUMEN
We have isolated and characterized a cDNA clone, termed OTK18, representing a novel human gene. The 3754 nucleotides of this clone contain an open reading frame of 2133 nucleotides. As the predicted 711-amino-acid protein contains 13 contiguous zinc-finger stuctures of the C2H2 type, it would be expected to function as a DNA-binding multi-finger protein. The 4.3-kb transcript was expressed in all adult human tissues examined by Northern analysis. The gene was assigned to chromosomal band 19q13.4 by fluorescence in situ hybridization.
Asunto(s)
Cromosomas Humanos Par 19 , Proteínas de Unión al ADN , Proteínas/genética , Factores de Transcripción , Dedos de Zinc/genética , Secuencia de Aminoácidos , Secuencia de Bases , Mapeo Cromosómico , Clonación Molecular , ADN Complementario , Humanos , Datos de Secuencia Molecular , Homología de Secuencia de AminoácidoRESUMEN
We have isolated a novel human cDNA clone that encodes a protein with 80% identity in amino acid sequence to DROER, the enhancer of the rudimentary gene in Drosophila melanogaster. The rudimentary gene product is thought to have significant enzymatic functions in the pyrimidine metabolic pathway, as well as a critical role in development of the wings. The human cDNA, termed ERH, consists of 797 nucleotides, which include an open reading frame of 312 nucleotides encoding 104 amino acids. The ERH gene was expressed in all normal human tissues examined. We assigned the ERH gene locus to chromosomal band 7q34 by fluorescence in situ hybridization.
Asunto(s)
Mapeo Cromosómico , ADN Complementario/genética , Drosophila melanogaster/genética , Elementos de Facilitación Genéticos , Genes de Insecto , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromosomas Humanos Par 7/genética , Clonación Molecular , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Homología de Secuencia de AminoácidoRESUMEN
We have isolated a novel human cDNA, designated RAB2L, that encodes a protein homologous to Ral guanine nucleotide dissociation stimulator (RalGDS). The predicted sequence of 122 amino acids revealed 39.5% identity with the C-terminal region of human RalGDS. As the C-terminal region of RalGDS is considered to interact with RAS, the product of this novel gene may also play some role in the ras-mediated signal transduction pathway. Northern analysis revealed that the gene RAB2L is expressed in all human tissues examined. We assigned this gene locus to chromosome band 6p21.3 by fluorescence in situ hybridization.
Asunto(s)
Cromosomas Humanos Par 6 , Proteínas de Unión al GTP/química , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP rab , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Bandeo Cromosómico , Mapeo Cromosómico , Clonación Molecular , Cósmidos , ADN Complementario/genética , Expresión Génica , Biblioteca de Genes , Factores de Intercambio de Guanina Nucleótido , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Factor de Intercambio de Guanina Nucleótido ral , Proteínas de Unión al GTP rapRESUMEN
We have isolated a novel human cDNA that encodes a protein highly homologous to NHP2, a nuclear protein of yeast, that is thought to have an essential physiological function for cell viability. The 1,493-bp cDNA sequence includes an open reading frame of 384 bp. This gene (NHP2L1) was expressed in all human tissues examined, and was localized to chromosome hand 12q24.3 by fluorescence in situ hybridization.
Asunto(s)
Mapeo Cromosómico , ADN Complementario/genética , Proteínas Fúngicas/genética , Proteínas Nucleares/genética , Ribonucleoproteínas Nucleares Pequeñas , Proteínas de Saccharomyces cerevisiae , Secuencia de Aminoácidos , Secuencia de Bases , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Saccharomyces cerevisiaeRESUMEN
The Wnt genes compose a large gene family encoding a group of secreted signaling molecules that have been implicated in oncogenesis and a number of developmental processes. We have isolated a full-length human cDNA clone that we consider to be a novel member of the Wnt gene family. The gene (WNT7A) encodes a deduced 349-amino-acid peptide with 98% identity in amino acid sequence to murine Wnt7a. Expression of this gene is restricted to certain tissues: placenta, kidney, testis, uterus, fetal lung, and fetal and adult brain. Furthermore, we have isolated a genomic clone of WNT7A and mapped it to chromosome 3p25 by fluorescent in situ hybridization.
Asunto(s)
Mapeo Cromosómico , Cromosomas Humanos Par 3 , Proteínas/genética , Proteínas Proto-Oncogénicas , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Clonación Molecular , ADN Complementario/genética , Femenino , Expresión Génica/genética , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Proteínas WntRESUMEN
Colligin, a collagen-binding glycoprotein localized to endoplasmic reticulum, belongs to the serpin superfamily. Colligin has been identified and cloned in a number of species including mouse, rat, chicken, and human. We have isolated and characterized a full-length human cDNA clone that encodes a 418-amino-acid peptide that is highly homologous (97% identity) to the previously reported human colligin gene, and have named this new member of the colligin family human colligin-2. The expression of the colligin-2 gene (CBP2) is ubiquitous among all normal human tissues except for brain and peripheral leukocytes. We have mapped this novel gene to chromosomes 11q13.5 by fluorescent in situ hybridization (FISH) using a cosmid clone containing the entire coding sequence and the 3' non-coding sequence.
Asunto(s)
Proteínas Portadoras/genética , Secuencia de Aminoácidos , Secuencia de Bases , Células Clonales , Glicoproteínas , Humanos , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Homología de Secuencia de AminoácidoRESUMEN
Fukuyama-type congenital muscular dystrophy (FCMD), the second most common form of childhood muscular dystrophy in Japan, is an autosomal recessive severe muscular dystrophy associated with an anomaly of the brain. After our initial mapping of the FCMD locus to chromosome 9q31-33, we further defined the locus within a region of approximately 5 cM between loci D9S127 and CA246, by homozygosity mapping in patients born to consanguineous marriages and by recombination analyses in other families. We also found evidence for strong linkage disequilibrium between FCMD and a polymorphic microsatellite marker, mfd220, which showed no recombination and a lod score of (Z) 17.49. A "111-bp" allele for the mfd220 locus was observed in 22 (34%) of 64 FCMD chromosomes, but it was present in only 1 of 120 normal chromosomes. This allelic association with FCMD was highly significant (chi 2 = 50.7; P < .0001). Hence, we suspect that the FCMD gene could lie within a few hundred kilobases of the mfd220 locus.
Asunto(s)
Mapeo Cromosómico , Desequilibrio de Ligamiento/genética , Distrofias Musculares/genética , ADN/análisis , Femenino , Genotipo , Humanos , Escala de Lod , Masculino , Distrofias Musculares/congénito , Recombinación Genética/genéticaRESUMEN
To isolate a putative tumor suppressor gene(s), we have constructed a physical map and a detailed deletion map of chromosome region 8p21.3-p22, where loss of heterozygosity (LOH) has been frequently seen in human hepatocellular carcinomas (HCC), colorectal cancers (CRC), and non-small cell lung cancers (NSCLC). The smallest commonly deleted region at 8p21.3-p22 in HCC and CRC was between the loci defined by C18-245 and C18-2644; in NSCLC, a region between C18-1051 and C18-2644 was commonly deleted. A contiguous physical map of 12 cosmid markers in the 8p21.3-p22 region was constructed by means of multi-color fluorescence in situ hybridization (FISH) and pulsed-field gel electrophoresis (PFGE). On the basis of this physical map, which spans roughly 3.1 Mb, the estimated sizes of the commonly deleted regions were at most 1.2 Mb in HCC and CRC and 0.6 Mb in NSCLC. As four of the 12 physically ordered markers are located within the 0.6 Mb region commonly deleted in all three tumor types, nearly one fourth to one fifth of the target region has already been covered with cosmid inserts.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/genética , Deleción Cromosómica , Cromosomas Humanos Par 8 , Neoplasias Colorrectales/genética , Neoplasias Hepáticas/genética , Neoplasias Pulmonares/genética , Animales , Autorradiografía , Mapeo Cromosómico , Electroforesis en Gel de Campo Pulsado , Genes Supresores de Tumor , Heterocigoto , Humanos , Células Híbridas , Hibridación Fluorescente in Situ , Ratones , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
We have determined the chromosomal localizations of newly isolated cosmids by fluorescent in situ hybridization (FISH) on prometaphase R-banded chromosomes and have constructed a high-resolution cytogenetic map for human chromosome 8 with 416 cosmid markers, including 328 new markers and 88 reported previously. Of the 416 markers, 229 were mapped to the long arm of chromosome 8, 181 to the short arm, and 6 to the centromere. Although the clones were scattered throughout the chromosome, they were concentrated in R-positive bands. Since the estimated physical length of chromosome 8 is 135 Mb, the overall average distance between loci is 320 kb, but the average separation of loci on R-positive bands is nearly 130-200 kb. This cytogenetic map will serve as a resource for efforts to characterize chromosomal and molecular aberrations involved in cancers, to clone genes associated with hereditary diseases, and to construct a detailed physical map of large electrophoretic fragments and/or contiguous cosmids and yeast artificial chromosomes.
Asunto(s)
Cromosomas Humanos Par 8 , Cósmidos , Marcadores Genéticos , Animales , Mapeo Cromosómico , Humanos , Células Híbridas , Hibridación Fluorescente in Situ , RatonesRESUMEN
Five cosmid clones, isolated by procedures to screen genomic libraries for homologous variants of the human prohibitin gene (PHB), were analyzed to determine their genomic structures. Four of these (PHBP1-4) were found to be processed pseudogenes, each located on a different chromosome from their counterparts on chromosome 17q21. The DNA sequence of one clone (PHBP1, on chromosome 6q25) shared a 91.3% identity at the nucleotide level with the cDNA of functional prohibitin. A large number of human tumors of the breast, ovary, liver, and lung were examined for somatic mutations in the PHB gene. Although mutations were observed in a few sporadic breast cancers, none were identified in any of the other cancers.
Asunto(s)
Genes Supresores de Tumor , Familia de Multigenes , Neoplasias/genética , Proteínas/genética , Proteínas Represoras , Secuencia de Bases , Neoplasias de la Mama/genética , Cromosomas Humanos Par 17 , Cromosomas Humanos Par 6 , Clonación Molecular , Cósmidos , ADN Complementario/genética , Femenino , Humanos , Hibridación Fluorescente in Situ , Neoplasias Hepáticas/genética , Neoplasias Pulmonares/genética , Datos de Secuencia Molecular , Mutación , Neoplasias Ováricas/genética , Prohibitinas , SeudogenesRESUMEN
As a critical step toward cloning the breakpoint of the (3;8) translocation observed in familial renal cell carcinoma and a common fragile site on chromosome 3p (FRA3B), we have characterized the 3p14 region containing the breakpoint and FRA3B by fluorescence in situ hybridization (FISH), pulsed-field gel electrophoresis (PFGE), and genetic linkage analysis. Of 23 cosmids mapped by FISH, 14 cosmids were distal to the breakpoint of t(3;8) and 9 were proximal. Analyses of FRA3B by FISH were identical to those for the (3;8) breakpoint and indicated that the breakpoint of t(3;8) occurred at or very close to the fragile site. We have also constructed a genetic linkage map and a preliminary long-range restriction map using PFGE of the 3p14 region. The linkage results were consistent with the physical data. The combined results of FISH, PFGE, and linkage analysis establish a basis for further experiments to clone the (3;8) breakpoint and FRA3B.
Asunto(s)
Carcinoma de Células Renales/genética , Fragilidad Cromosómica , Cromosomas Humanos Par 3 , Cromosomas Humanos Par 8 , Neoplasias Renales/genética , Translocación Genética , Animales , Southern Blotting , Sitios Frágiles del Cromosoma , Mapeo Cromosómico , Cósmidos , Electroforesis en Gel de Campo Pulsado , Ligamiento Genético , Humanos , Células Híbridas , Hibridación Fluorescente in Situ , RatonesRESUMEN
Relationship between prolonged life span and changes of serum IAP and albumin induced by the therapy of lentinan plus tegafur were analysed on 43 cases with inoperable and recurrent gastric cancer. Antitumor effect was observed only in one case (2%). Other clinical effects such as improvement of performance status (PS), appetite or pain were observed in 18 cases (42%). Decrease of serum IAP was observed in 25 cases (58%) and increase of albumin was observed in 20 cases (47%). The changes of these two factors seemed to be reversely correlated. Among 30 cases which didn't show decrease of albumin, we found no increase of serum-IAP in 22 cases (72%). In the cases which showed decrease of serum IAP from abnormally high level (more than 500 mu/ml) and increase of albumin from abnormally low level (less than 3.5g/dl), prolonged life span was observed by comparison with the other cases. The cases with any clinical effect contained 78% of the cases without increase of serum IAP, and 72% of the cases without decrease of albumin. These results suggested that life prolongation effect or improvement of clinical symptoms by our therapy was closely related to the change of these serum factors.