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Biomedical subjects

S Surrey

Publications and source records attributed to S Surrey.

At least 109 records · Page 6Linked to original sources
PubMed1987

Transcriptional activity of the genes for apoproteins A-I and E in neonatal rat liver.

The expression of the genes for apoproteins A-I and E (apoA-I, apoE) in fetal and neonatal rat livers was quantitated by measurement of steady-state levels of mRNA and by assays of relative transcriptional activity using the nuclear 'run-on' technique. From the last day of gestation (day -1) to day 5 after birth (day 5), for apoE there was a 2.2-fold increase in relative transcriptional activity and a 2.6-fold increase in steady-state mRNA. For apoA-I, from day -1 to 3 there was a 1.25-fold increase in both transcription and steady-state mRNA levels. We conclude that the increase in steady-state mRNA for the apoE gene which occurs during liver development in the rat is facilitated primarily by transcriptional control.

Animals↗
PubMed1987

The xerocytosis of Hb SC disease.

Patients with Hb SC disease were found to have microcytic and hyperchromic red cell indices despite mild reticulocytosis. Iron deficiency anemia was ruled out by the finding of normal serum ferritin levels. In order to determine whether the microcytosis was due to coexistent alpha-thalassemia, restriction endonuclease mapping was performed on genomic DNA extracted from peripheral blood leukocytes. Patients with Hb SC disease had microcytic indices despite the presence of a full complement of four alpha-genes (alpha alpha/alpha alpha), suggesting that the microcytosis may be due to cellular dehydration (or xerocytosis), since the mean corpuscular hemoglobin concentration in Hb SC disease patients was significantly higher than in controls. This possibility was investigated further by the determination of RBC cation content. RBC Na levels were similar in SC and normal red cells. Hb SC RBCs, however, had significantly reduced K levels. These findings show that RBC cation content, and thus cell water, is decreased in Hb SC disease. The decreased RBC K level in the presence of normal cellular Na concentration suggests selective K loss that is not due to inhibition of the Na K pump. Ouabain-insensitive K+ efflux was increased to four times normal in SC cells. Cell dehydration was confirmed by the demonstration of increased high-density RBCs on discontinuous Stractan density gradients and by osmotic gradient ektacytometry. Cellular dehydration and its sequelae were worse in CC erythrocytes and milder in AC cells than in Hb SC red cells. Taken together, these data indicate that in Hb SC disease the RBCs are severely dehydrated and typically microcytic and hyperchromic. Hb SC RBCs seem to be dehydrated due to selective K loss. These findings suggest a functional interrelationship between Hb SC, the red cell membrane, and cation regulation.

Adenosine Triphosphatases↗
PubMed1987

Cloning and characterization of platelet factor 4 cDNA derived from a human erythroleukemic cell line.

We report the isolation of a platelet factor 4 (PF4) cDNA clone from a lambda gt11 expression cDNA library which was derived from a human erythroleukemic (HEL) cell line. The sequence of the DNA insert includes the 3'-untranslated region, the entire amino acid coding region for the mature PF4 protein, and a 5' region containing coding information for an additional 18 amino acids. In addition, supplemental genomic DNA sequencing shows that the full-length leader sequence is 30 amino acids long plus an initial methionine and codes for a hydrophobic signal-like sequence which is probably involved in transmembrane transport. A single species mRNA of approximately 800 nucleotides was detected on blots of HEL cell poly(A) + RNA using a labeled PF4 cDNA probe. The human PF4 leader sequence shares some DNA, but no amino acid, homology with the 15 amino acids at the N-terminus of mature bovine PF4, suggesting rapid divergence in this region of PF4 between these two species. Sequence comparison of the coding regions of mature PF4 and gamma IP-10, a protein induced in a variety of cells following treatment with gamma-interferon, shows a corrected divergence of 76%. The divergence of a common ancestor protein into PF4 and gamma IP-10 may have accompanied the development of sophisticated immune and coagulation systems in vertebrates. The availability of cDNA and genomic DNA information for these genes in other species will be useful in studying the evolution of the coagulation and immune systems.

Amino Acid Sequence↗
PubMed1985

Functional analysis of a beta-globin gene containing a TATA box mutation from a Kurdish Jew with beta thalassemia.

We recently reported a TATA box mutation (ATAAAA to ATACAA) in a cloned beta-globin gene from a Kurdish Jew with homozygous beta thalassemia (Poncz, M., Ballantine, M., Solowiejczyk, D., Barak, I., Schwartz, E., and Surrey, S. (1982) J. Biol. Chem. 257, 5994-5996). We have now introduced this gene into HeLa cells after CaPO4 precipitation of the DNA and studied expression by analyzing globin-gene transcripts with a novel S1 nuclease mapping assay. Quantitative and qualitative comparison with the normal beta-globin gene revealed a promoter-down phenotype in the TATA box mutant, with normal RNA processing, and a normal start site for initiation of the primary transcript. Decreased transcriptional efficiency was confirmed directly by analysis of run-off transcripts using assays in vitro. The patient's phenotype of beta thalassemia major is probably the result of two different mutations since haplotype analysis of the beta-like globin gene clusters in genomic DNA from this patient shows heterozygosity for the Mediterranean-type haplotypes I and VII, with the TATA box mutation on a haplotype I chromosomal background.

Base Sequence↗
PubMed1984

Hemoglobin I mutation encoded at both alpha-globin loci on the same chromosome: concerted evolution in the human genome.

Genetic analysis of an individual expressing an unexpectedly high level of hemoglobin I, an alpha-globin structural mutant, reveals that the mutation is present at both the alpha 1- and the alpha 2-globin gene loci. Kindred analysis confirms that the two affected genes are located in cis. The most likely explanation for this finding is that a recent conversion event occurred within the human alpha-globin gene cluster.

Base Sequence↗
PubMed1984

Frequent occurrence of a zeta-globin-region deletion in American blacks accounts for a previously-described restriction site polymorphism.

We describe an 11.3 kb deletion within the zeta-globin gene region which was found in 2 of 16 American Black neonates. The deletion is consistent with an unequal crossover between the homologous zeta and psi zeta gene regions, resulting in a shortened Eco RI fragment (17.2 kb) containing a single zeta and the psi alpha, alpha 2, and alpha 1 genes. An earlier report of zeta-region crossovers (1) may have overestimated the size of this fragment by 2 kb. This finding led us to re-evaluate another report which concluded that a 17 kb Eco RI fragment found in 10% of the Black population was the result of a restriction site polymorphism (2). Our data do not substantiate this conclusion and we propose that the high frequency of this shortened fragment is due instead to an unequal crossover. Possible clinical consequences of this deletional event are also discussed.

Black People↗
PubMed1984

Detection of a novel DNA polymorphism in the beta-globin gene cluster.

Analysis of DNA from the beta-globin gene cluster in an Albanian family identified a novel RsaI site approximately 550 base pairs 5' to the beta-globin gene. In this family, two chromosomes carrying otherwise identical beta-globin haplotypes were found to differ at the RsaI site. Population screening demonstrated the presence and absence of the site in DNA from individuals of northern European, Mediterranean, Middle Eastern, Southeast Asian, African, and Asian Indian descent, indicating that this site is a DNA polymorphism common in many ethnic groups. The polymorphism is also present in DNA from individuals carrying different beta-globin alleles. Additional nucleotide sequence changes identified in an RsaI (+) genomic clone in the region immediately 3' to the RsaI site suggest a mechanism for the randomization of the site with respect to haplotype.

Alleles↗
PubMed1984

The silent carrier allele: beta thalassemia without a mutation in the beta-globin gene or its immediate flanking regions.

A molecular genetic analysis has been performed using as subjects an Albanian family in which the father is a silent carrier, the mother has high Hb A2-beta thalassemia trait, and both children have beta thalassemia. Nucleotide sequence analysis of the daughter's paternal beta-globin gene and its flanking regions failed to reveal any base changes of known functional significance. When introduced into HeLa cells the gene was expressed at normal levels with proper processing of RNA. Haplotype analysis revealed that the affected son and daughter inherited different epsilon gamma delta beta-globin gene clusters from the father. The silent carrier allele is not due to a mutation within the beta-globin structural gene or its flanking regions and as such represents a novel form of beta+ thalassemia.

Alleles↗
PubMed1983

Nucleotide sequence analysis of the delta beta-globin gene region in humans.

The continuous DNA sequence of a 16.5-kilobase pair region encompassing the linked delta beta-globin gene cluster in humans is presented with a detailed restriction endonuclease map. There are 38 differences (0.5%) in comparison with published sequence data, corrected for errors in sequencing, resulting in polymorphic rates of 0.2% in exons and 0.76% in 5'-gene flanking regions. Fifteen changes result in the generation or elimination of restriction sites which may be useful in linkage disequilibrium studies. Two pairs of inverted Alu repeats, a pyrimidine-rich region 5' to delta, and (TG)n, (Pu/Py)n, and (ATTTT)n tracts 5' to beta are described. Dinucleotide frequencies and deviation from expected values approximated those found in total human genomic DNA. Regions of less than 50% A + T content were found associated with Alu sequences, a 150-base pair region immediately 5' to the beta gene, exon regions from both genes, and an area 3' to the beta gene. These regions also contained significantly lower than expected CpG levels compared to other regions, suggesting a possible relationship between DNA organizational patterns and functionally important regions. In addition, strand asymmetries in base composition in this region differ from those associated with the fetal globin genes.

Base Composition↗
PubMed1982

beta-Thalassemia in a Kurdish Jew. Single base changes in the T-A-T-A box.

We recently described a "non-random" sequencing procedure for DNA inserts in bacteriophage M13 using Bal 3 nuclease and the dideoxy chain termination method (Poncz, M., Solowiejczyk, D., Ballantine, M., Schwartz, E., and Surrey, S. (1982) Proc. Natl. Acad. Sci. U. S. A., in press). Using this procedure, we have determined the nucleotide sequence of a cloned human beta-globin gene from a Kurdish Jew with beta +-thalassemia major. Comparison with the previously reported human beta-globin gene sequences (1-3) reveals a change in the "T-A-T-A" box. This region 5' to the capping site was previously demonstrated to be critical for the proper transcription in vitro of several different eukaryotic genes (4-7). This is the first report of a T-A-T-A box modification found in association with a spontaneously occurring human genetic disorder. In addition to this mutation, other base changes, an insertion, and a deletion in the cloned gene were found in the 5' and 3' flanking regions.

Base Sequence↗
PubMed1982

"Nonrandom" DNA sequence analysis in bacteriophage M13 by the dideoxy chain-termination method.

We describe a rapid "nonrandom" DNA sequence analysis procedure that facilitates the nucleotide sequence determination of large contiguous regions of DNA. The method consists of cloning a restriction endonuclease fragment of interest into bacteriophage M13 followed by construction of a series of nuclease BAL-31 deletion mutants originating from a single site in M13 that is close to the DNA insert. Determination of the size of the deletion mutant is accomplished by hybridization to a complementary single-stranded probe derived from M13 containing that total insert followed by nuclease S1 treatment. Single-stranded M13-insert DNAs of progressively smaller sizes are isolated and analyzed by using a site-specific M13 DNA primer and the dideoxy chain-termination method. In this way, analysis of the DNA sequence proceeds from one end of the total insert to the other in a nonrandom fashion due to generation of a controlled overlapping set of deletion mutants.

Base Sequence↗
PubMed1982

Construction of human gene libraries from small amounts of peripheral blood: analysis of beta-like globin genes.

We describe a rapid procedure for constructing cloned human genomic libraries from small amounts of peripheral blood. High molecular weight DNA is isolated from 5-20 ml peripheral blood, partially cleaved with Eco R1, and 8-22 kb fragments are cloned using bacteriophage Charon 4A and suitable E. coli host. Using the approach we have isolated and characterized several non-alpha globin clones from a Kurdish Jew with homozygous beta thalassemia. The ability to isolate suitable amounts of high molecular weight DNA from peripheral blood provides a relatively simple means of constructing human gene libraries representing a variety of hemoglobin disorders.

Base Sequence↗
PubMed1982

The gamma-delta-beta-globin gene region in G gamma-beta +-hereditary persistence of fetal hemoglobin.

We report restriction endonuclease analysis of the gamma-delta-beta-globin gene region in a mother and child heterozygous for G gamma-beta +-hereditary persistence of fetal hemoglobin (HPFH). The affected chromosome in these persons directs the production of G gamma-chains and beta-chains but not A gamma-chains. DNA was digested with several restriction enzymes and was examined for gamma, delta, beta sequences by blot hybridization. Only normal digestion fragments were present. By sensitive methods, we were unable to detect a deletion in the entire gamma-delta-beta-globin gene region of the affected chromosome, indicating that in this family, G gamma-beta +-HPFH is not due to a large deletion.

Adult↗
PubMed1980

Linkage of alpha G-Philadelphia to alpha-thalassemia in African-Americans .

We have studied the inheritance of the alpha-chain hemoglobin variant Hb G-Philadelphia (alpha 2(68 Asn leads to Lys)Beta 2) in two African-American families. Expression of the alpha-globin loci was monitored by the percentage of Hb G in these individuals. The variant represented approximately 33% of the total adult hemoglobin in some and 50% in others. alpha-Globin gene fragments were analyzed by using restricton endonucleases that cleave outside (EcoRI), within (HindIII), and between (Bgl II) the normal duplicated alpha-globin loci (alpha alpha/alpha alpha). Individuals having 33% variant lack one functioning alpha gene (alpha G/alpha alpha); those with 50% variant lack two genes, one missing on each chromosome (alpha G/alpha). Inheritance of alpha G was therefore linked to that of a chromosome with only one functional alpha-globin gene locus. This locus is probably the result of a nonhomologous crossover. Our results also suggest equal expression of the alpha-globin loci in humans because the percentages of the variant could be explained solely on the basis of the total number of alpha genes present. The percentages of Hb G as well as other hematologic data all were consistent with the number of alpha-globin genes identified by restriction endonuclease mapping. Gene mapping yields a more precise determination of the number of alpha-globin genes than does study of globin synthesis.

Africa↗
PubMed1980

alpha Thalassemia and the expression of hemoglobin G-Philadelphia.

The findings presented for these two families help to explain the inheritance of alpha thalassemia, alpha-chain variants, and the relative expression of alpha genes. An 18.0-kb EcoRI fragment contains only one functional alpha gene, whereas a 20.5-kb fragment contains two. Individuals homozygous for the 18.0-kb EcoRI fragment also lack the 4.1-kb HindIII fragment that normally connects the centers of the duplicated alpha genes. These findings are consistent with a deletion involving the 5' alpha-gene locus. Presence of alpha G-Philadelphia in both families was found in association with the 18.0-kb EcoRI fragment; this short fragment was also found in an individual with alpha A. Inheritance of alpha G-Philadelphia at one alpha-gene locus was therefore also linked to the inheritance of alpha thalassemia due to a deletion involving the second alpha gene. The high percentage (46% to 48%) of alpha G found in some family members was due to alpha-thalassemia trait, or deletion of two alpha genes (-alpha G/-alpha); others with levels of variant of 32% to 34% were shown to have three functional alpha genes (-alpha G/alpha alpha). The genetic expression of the four normal alpha genes therefore appears to be equal and furthermore implies the existence of separate independently functioning transcriptional units for each of these genes in humans. It would be interesting to analyze the alpha genes in Afro-Americans reported to have alpha G-Philadelphia in the 20% to 25% range to determine whether the inheritance of alpha G can be linked to a normal alpha gene.

Base Sequence↗
PubMed1980

Alpha-gene deletions in black newborn infants with Hb Bart's.

The presence of increased Hb Bart's (gamma 4) in cord blood is believed to be an indication of alpha-thalassemia. We have used restriction endonuclease nalyses of DNA to compare the number of alpha-genes with the percentage of Hb Bart's in 6 older children who had Hb Bart's at birth and 17 newborns. Four children with > 2% Hb Bart's had Eco R1 alpha-gene patterns and hematologic data consistent with the presence of two alpha-genes, one per chromosome. Of the remaining 19 children, all of whom had < 2% Hb Bart's, 8 had 3, while 11 had 4 alpha-genes. Three infants with Hb Bart's between 1% or less Hb Bart's. infants with 3 alpha-genes may therefore have elevated or normal levels of Hb Bart's at birth. DNA analysis is the definitive method for the determination of heterozygous alpha-thalassemia syndromes in newborns.

Black People↗