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J E Hixson

Publications and source records attributed to J E Hixson.

At least 73 records · Page 4Linked to original sources
PubMed1992

A DNA polymorphism for LCAT is associated with altered LCAT activity and high density lipoprotein size distributions in baboons.

A polymorphic Pvu II site was mapped to intron 5 of LCAT, the gene encoding baboon lecithin: cholesterol acyltransferase (LCAT). In a study of 83 baboons, heterozygous baboons (Pv1/Pv2) had significantly higher LCAT enzyme activity levels than did baboons homozygous for the more common allele (Pv1/Pv1). LCAT genotype explained 6% of the total variation in LCAT enzyme activity. To test for allelic effects on cholesterol metabolism, we compared serum concentrations of high density lipoprotein (HDL) cholesterol and apolipoprotein A-I (apo A-I). We also compared distributions of cholesterol and apo A-I among three HDL size classes (HDL1, HDL2, and HDL3). All measurements were obtained for each baboon after long-term feeding of a basal diet low in cholesterol and fat and again after 7 weeks on an atherogenic diet. Heterozygous baboons had significantly lower serum levels of total cholesterol than did homozygotes. In addition, we detected significant effects of LCAT genotype on size distributions of HDL cholesterol and apo A-I on both diets but did not detect any genotype-by-diet interaction. Heterozygotes had increased amounts of cholesterol and apo A-I in HDL3 particles and lower amounts of cholesterol and apo A-I in the larger HDL size classes by comparison with homozygotes. Overall, the LCAT polymorphism explained a significant proportion of total variation in cholesterol (4-10%) and apo A-I (13%) distributions on both diets. Thus, the results indicate that the LCAT polymorphism is associated with significant differences in LCAT enzyme activity and with alterations in HDL compositions.

Animals↗
PubMed1992

Apo B insertion/deletion polymorphisms are associated with atherosclerosis in young black but not young white males. Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Research Group.

Investigators in eight communities collected aortas, right coronary arteries, blood and liver samples, and associated information from 872 young males, aged 15-34 years, who died of external causes. Pathologists graded the arteries for atherosclerotic lesions, and a central laboratory measured lipoprotein cholesterol concentrations. Apolipoprotein (apo) B sequences were amplified in hepatic DNA samples to determine genotypes for length polymorphisms in the signal peptide of apo B. In addition to the insertion (ins) allele (27-amino acid signal peptide) and the deletion (del) allele (24 amino acids), we detected a rare allele (ins*) in whites with an in-frame insertion of two Leu codons in a region that normally contains six Leu codons. The frequency for the apo B del allele was lower in blacks than in whites (p less than 0.0001). In blacks, homozygotes for the ins allele had the lowest levels of serum cholesterol and very low plus low density lipoprotein cholesterol (VLDL + LDL-C), homozygotes for the del allele had the highest levels, and heterozygotes had intermediate levels (p = 0.0509 for cholesterol, p = 0.0530 for VLDL + LDL-C), but no differences were found in whites. In blacks, homozygotes for the ins allele had the least involvement of the thoracic and the abdominal aorta with lesions, homozygotes for the del allele had the greatest involvement, and heterozygotes had intermediate involvement (p = 0.0328 for thoracic aorta, p = 0.0104 for abdominal aorta), but no differences were found in whites.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

Restriction isotyping of human apolipoprotein A-IV: rapid typing of known isoforms and detection of a new isoform that deletes a conserved repeat.

Genetic polymorphisms of apolipoprotein A-IV (apoA-IV) have been detected by isoelectric focusing of serum proteins. Because genetic variation in apoA-IV has significant effects on lipid risk factors, we used restriction enzyme isoform genotyping (restriction isotyping) to determine apoA-IV isoform genotypes at the DNA level for a large population (n = 509). In contrast to isoelectric focusing methods, restriction isotyping relies on nucleotide differences, enabling unambiguous typing of known isoforms and detection of new alleles that mimic other isoforms with shared charge properties. To determine genotypes for the common A-IV-1 isoform (Gln at aa position 360) and A-IV-2 isoform (360His), we used a mismatched primer for polymerase chain reaction (PCR) to introduce a restriction site (PvuII) that distinguishes each isoform. Using a portion of the same PCR reaction, we used HinfI to distinguish isoforms with Thr at position 347 (347Thr) versus Ser (347Ser). In surveys for these common genotypes, we detected heterozygotes for an allele with an insertion of 12 bp. Nucleotide sequencing showed that this allele is identical to the A-IV-0 isoform that inserts a hydrophilic repeat (Glu Gln Gln Gln) in a conserved region near the carboxy terminus. In addition, we discovered a new allele with a 12 bp deletion that removes a repeat (Glu Gln Gln Gln) from the same region. Nucleotide sequencing showed that this allele removes an acidic charge relative to A-IV-1, so we have named this isoform A-IV-2*. This isoform has not been discovered at the protein level, perhaps due to shared charge properties with A-IV-2 isoforms.

Adolescent↗
PubMed1990

Lipid phenotypes, apolipoprotein genotypes and cardiovascular risk in nonagenarians.

Despite great interest in the role of lipids in overall and disease-free survival, virtually no information is available on the lipids, lipoproteins and apolipoproteins of persons over 90 years of age. Furthermore, the genetic underpinnings of atherosclerosis and the particular genetic factors responsible for protection against coronary artery disease remain speculative. In Bloomfield, Nebraska, we studied 41 nonagenarians (10 males, 31 females), with a mean age of 92.7 years, in whom lipids, lipoproteins, apolipoproteins and restriction fragment length polymorphisms (RFLPs) of genes for apolipoprotein B (apo B), aop AI and apo CIII were assessed. Nearly complete historical, physical and laboratory data were obtained on 39 subjects. The mean diastolic and systolic blood pressures for this group were nonhypertensive, body mass indices (weight/height2) had a mean of 23.9 and triceps skinfold thickness measurements an overall mean of 14.8 mm. The mean total serum cholesterol was 5.42 mmol/l. HDL-cholesterol levels in females persisted to be higher when compared to males (P less than 0.013). The allele frequencies for apo AI (MspI and PstI), apo CIII (Sst) and apo B (XbaI) gene RFLPs were typical for larger population studies. In these preliminary studies, we did not identify a distinctive phenotype, genotype, or phenotype-genotype relationship. Diversity of cardiovascular risk was the hallmark of these nonagenarians.

Aged↗
PubMed1990

Restriction isotyping of human apolipoprotein E by gene amplification and cleavage with HhaI.

We have used restriction isotyping (restriction enzyme isoform genotyping) for rapid typing of common apolipoprotein E isoforms (E2, E3, E4). ApoE restriction isotyping used oligonucleotides to amplify apolipoprotein E gene sequences containing amino acid positions 112 and 158. The amplification products were digested with HhaI and subjected to electrophoresis on polyacrylamide gels. Each of the isoforms was distinguished by a unique combination of HhaI fragment sizes that enabled unambiguous typing of all homozygotic and heterozygotic combinations. HhaI cleaves at GCGC encoding 112arg (E4) and 158arg (E3, E4), but does not cut at GTGC encoding 112cys (E2, E3) and 158cys (E2).

Amino Acid Sequence↗
PubMed1989

Apolipoprotein(a) (Apo(a)) glycoprotein isoforms result from size differences in Apo(a) mRNA in baboons.

Like humans, baboons possess serum lipoprotein(a) that contains apolipoprotein(a) (apo(a], and baboon apo(a) also occurs in distinguishable glycoprotein isoforms. To investigate the molecular basis for isoform size variation, we isolated hepatic RNA from baboons possessing different apo(a) isoforms for Northern blot analyses with rhesus apo(a) cDNA probes. In a survey of 22 baboons, we detected a variety of sizes of apo(a) transcripts (5.2-11.2 kilobases) that corresponded with relative mobilities and numbers of serum apo(a) isoforms. In unrelated baboons possessing apo(a) isoforms of similar mobilities, we detected apo(a) transcripts with similar mobilities. In related baboons possessing apo(a) isoforms that were identical by descent, apo(a) transcripts were also identical in size. Using data from 22 baboons included in this study, we compared the sizes of 20 apo(a) isoforms and corresponding transcripts. Linear regression analysis established a highly significant correlation (p less than 0.001) between estimated apo(a) transcript size and glycoprotein mass (r2 = 0.996). From these results, we conclude that apo(a) glycoprotein isoforms are due to structural differences in apo(a) transcripts. However, we also noted exceptions to correspondence between apo(a) transcripts and isoforms that suggest the action of additional post-transcriptional control of serum apo(a) levels.

Alleles↗
PubMed1989

Alpha-myosin heavy chain cDNA structure and gene expression in adult, fetal, and premature baboon myocardium.

To examine cardiac myosin gene structure and expression in a non-human primate model for human heart development and disease, we have constructed a cDNA library from baboon atrium and used baboon beta-myosin heavy chain (beta-MHC)* cDNA probes to isolate atrial MHC clones. The nucleotide sequence of one such clone, lambda BMHC alpha 3, contains sequences that encode part of the light meromyosin region (LMM) and the 3' untranslated region of the baboon alpha-MHC. To study cardiac MHC gene transcription, we constructed probes from the baboon alpha-MHC cDNA for S1 nuclease analyses of RNA from atria and ventricles. To examine translational regulation of cardiac MHC gene expression, we used monoclonal antibodies (MAb) against specific alpha- and beta-MHC epitopes for Western blot analyses. In atria and ventricles from adult baboons, we detected predominantly alpha- and beta-MHC gene transcripts, respectively. In ventricles from fetal baboons at two stages of development (140 and 160 days gestation), we also detected predominantly beta-MHC gene transcripts and isoforms. To investigate changes induced by parturition, we obtained ventricles from baboons that were prematurely delivered at 140 days gestation and supported for 10 days in an extrauterine environment. In contrast to adult and fetal patterns, we observed an increase in alpha-MHC transcripts and isoforms in ventricles of premature baboons. Because alpha-MHC gene expression is increased in premature baboons (total age of 150 days) compared to their older 160 day fetal counterparts, the induction of ventricular alpha-MHC synthesis must have resulted from factor(s) associated with parturition or prolonged mechanical ventilation rather than at predetermined stages of gestational development.

Amino Acid Sequence↗
PubMed1988

The baboon gene for apolipoprotein A-I: characterization of a cDNA clone and identification of DNA polymorphisms for genetic studies of cholesterol metabolism.

We have isolated and sequenced a baboon apolipoprotein A-I (ApoA-I) clone from a liver cDNA library using a human cDNA hybridization probe. This baboon cDNA contains the entire ApoA-I coding region (801 bp, 267 aa), a 3' untranslated region, and a poly(A)tail. Among comparisons with apoAI sequences from other species, the baboon cDNA is most similar to that of the cynomolgus macaque (99.2% homologous) and least similar to the rat sequence (72.6% homologous). A high frequency of nonsynonymous substitutions are observed by alignment of baboon and human apoAI cDNAs, but comparisons of hydrophilicity profiles show that protein structure is conserved by substitutions of aa with similar properties. A polymorphic PstI cleavage site was identified by Southern blot analysis and subsequently mapped to the 5' end of the baboon apoAI gene. To identify effects of apoAI allelic variation on cholesterol metabolism, we used immunoblotting to compare the distributions of ApoA-I among lipoprotein size classes in baboons from each genotype under basal and atherogenic diets. We observed an increase of ApoA-I in high density lipoprotein (size class 1) particles after atherogenic diets in homozygotes for one allele, as compared to slight decreases in the other genotypes.

Amino Acid Sequence↗
PubMed1988

Comparisons of ape and human sequences that regulate mitochondrial DNA transcription and D-loop DNA synthesis.

The mitochondrial DNA (mtDNA) control regions for common chimpanzee, pygmy chimpanzee and gorilla were sequenced and the lengths and termini of their D-loop DNA's characterized. In these and all other species for which there are data, 5' termini map to sequences that contain the trinucleotide YAY. 3' termini are 25-51 nucleotides downstream from a sequence that is moderately conserved among vertebrates. Substitutions were greater than 1.5 times more frequent in the control region than in regions encoding structural genes. Additions and deletions were also frequent, especially in gorilla. Sequences of promoters and of two of four transcription factor binding sites were highly conserved. Comparisons of sequence similarity and transition/transversion ratios suggest that human and chimpanzees may be more closely related to each other than either is to gorilla, if substitution rates are approximately equal among these species.

Animals↗
PubMed1988

The baboon beta-myosin heavy-chain gene: construction and characterization of cDNA clones and gene expression in cardiac tissues.

We have constructed a cDNA library from baboon ventricle and have used a rabbit beta-myosin heavy chain (beta-MHC) cDNA probe to isolate cross-hybridizing clones. The nucleotide sequence of one such clone, lambda BMHC beta 14, contains a portion of the coding region of the light meromyosin (LMM) region and the 3'-untranslated region of the baboon ventricular MHC. This cDNA clone is identified as containing beta-MHC sequences on the basis of similarity with the 3'-untranslated regions of beta-MHC genes from man (96% homologous) and rat (71% homologous), and dissimilarity with the 3'-untranslated region of the rat alpha-MHC gene (25% homologous). Alignment and comparison of the baboon cDNA nucleotide sequence with a human cDNA sequence reveal two amino acid substitutions in the LMM region that cause differences in their hydrophilicity profiles. These differences may alter MHC functions such as filament assembly. We have used baboon beta-MHC cDNA clones to construct probes for S1 nuclease protection studies to detect and to distinguish cardiac MHC gene transcripts in baboon ventricle, atrium, and diaphragm. As in human tissues, beta-MHC gene transcripts are detected in RNA from baboon ventricle and diaphragm. In baboon atrium, we detect beta-MHC gene transcripts as well as transcripts that may represent expression of the alpha-MHC gene. This study represents the first examination of cardiac MHC genes and gene expression in tissues from a large mammal that is closely related to man.

Amino Acid Sequence↗
PubMed1988

The baboon apolipoprotein E gene: structure, expression, and linkage with the gene for apolipoprotein C-1.

To develop the baboon model for molecular genetic studies of atherosclerosis, we have cloned and sequenced the baboon apolipoprotein E (apo E) gene. The baboon apo E gene encodes the E4 isoform with respect to specific amino acid positions, suggesting that the common epsilon 3 allele is not the primal human allele. Rather than accumulating predominantly synonymous nucleotide changes, 50% of substitutions in human and baboon apo E gene coding regions cause amino acid substitutions. However, comparisons of these apo E proteins show conservation of amphipathic helices required for apo E--lipid interactions. The human and baboon apo E genes have diverged less extensively than those from rat and mouse, providing further evidence for a slowing of molecular evolution in primate species. The baboon and rhesus monkey apo E genes (intron 2) contain two Alu repeats that are absent in the human gene, indicating insertion after the divergence of human and cercopithecine lineages, but before the baboon/rhesus divergence. S1 nuclease studies show that transcription of the baboon apo E gene starts at two different positions, one of which corresponds to the human gene start site. To examine linkage of apolipoprotein genes in the baboon genome, we have used a human cDNA probe to detect apo C-I gene sequences approximately 4 kb from the 3' end of the baboon apo E gene.

Amino Acid Sequence↗
PubMed1987

DNA markers in primate models of human disease.

Nonhuman primates are particularly useful as animal models for common human diseases in which both genetic and environmental factors play important roles. The recent development of DNA markers (restriction fragment length polymorphisms, RFLPs) greatly increases the power of linkage analysis to detect major genes that affect quantitative phenotypes, including those related to diseases. This paper summarizes a strategy for using RFLPs in linkage analysis of baboon pedigrees to identify genes that control lipoprotein phenotype, which in turn is predictive of susceptibility to atherosclerosis. This strategy also can be applied to other common human diseases for which nonhuman primate models exist.

Animals↗
PubMed1986

Both the conserved stem-loop and divergent 5'-flanking sequences are required for initiation at the human mitochondrial origin of light-strand DNA replication.

Mammalian mitochondrial DNAs contain a conserved origin of light-strand replication that supports accurate initiation of DNA synthesis in vitro. This provides an opportunity to examine the sequence requirements for initiation through in vitro analysis of a series of deleted and mutagenized DNA templates. These assays use enzymes isolated from human mitochondria and single-stranded DNA templates containing deletions or substitutions in the known origin region. The data indicate that accurate and efficient light-strand replication in vitro requires the previously identified stem-loop structure located within a tRNA cluster. In addition, the template sequence 3'-GGCCG-5', located immediately adjacent to the stem, is necessary for efficient replication. This sequence, the complement of which encodes the 3' end of tRNACys, may be the site of transition from RNA primer synthesis to DNA synthesis. Surprisingly, substitutions within a region located in the loop of this origin do not reduce levels of replication.

Base Sequence↗
PubMed1986

A comparison of the small ribosomal RNA genes from the mitochondrial DNA of the great apes and humans: sequence, structure, evolution, and phylogenetic implications.

Restriction endonuclease fragments produced by EcoRI/AvaI or KpnI digestion and containing the small (12S) ribosomal RNA (rRNA) genes from the mitochondrial DNAs (mtDNAs) of the common chimpanzee, pygmy chimpanzee, gorilla, and orangutan were inserted into the plasmids pBR322 or pADD1. After species verification the inserted fragments were digested with SauIIIA, subcloned into M13mp7 vectors, and sequenced. The small rRNA gene sequences were compared with each other and with the published human sequence (Anderson et al. 1981). Substitutions were detected at 118 of the 955 nucleotide positions compared. Pairwise, the sequence differences ranged from 1% (between the chimpanzee species) to 9% (comparisons involving the orangutan); the proportion that were transitions ranged from 87% to 100%. Deletions and/or additions were noted at seven locations. With respect to evolutionary sequence lability, kinetic analysis indicated the presence of at least two classes of nucleotide positions; the more labile class occurs in sequences thought to form self-complementary duplexes (stems) in the mature rRNA. The high frequency of compensating substitutions, which maintain base-pairing within these sequences, corroborates their inferred structure. Phylogenetic inferences drawn from the sequence comparisons support the notion of an approximately equidistant relationship among chimpanzees, gorilla, and man, with the orangutan much less closely related. However, inference from a shared deletion suggests that the gorilla and the chimpanzees may be more closely related to one another than they are to man.

Animals↗
PubMed1986

Minor transcription initiation events indicate that both human mitochondrial promoters function bidirectionally.

Human mitochondrial DNA is transcribed from two distinct, strand-specific promoters located in the displacement loop region of the genome. The transcriptional control sequences identified by deletion mapping and site-directed mutagenesis studies span short regions surrounding the initiation sites and bear no obvious sequence homology to any nuclear or procaryotic promoters. In vitro transcription analyses also revealed several minor initiation sites that are characterized by a pyrimidine-rich region followed by a purine-rich region, a feature that is shared by the two major promoters. In this paper, we report a new class of minor promoters in human mitochondrial DNA. These minor promoters were localized to the same duplex DNA sequences that direct major transcriptional events, but they had transcriptional polarity opposite to that of the major promoters. Furthermore, nucleotide changes that affected the major form of transcription similarly affected transcription in the opposite direction. For one of these minor promoters, a corresponding in vivo RNA species initiating from the same site was identified. These observations indicate that the major transcriptional promoters in human mitochondria can function bidirectionally both in vivo and in vitro.

Base Sequence↗