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J M Weber

Publications and source records attributed to J M Weber.

At least 19 recordsLinked to original sources
PubMed1992

Pathways for oxidative fuel provision to working muscles: ecological consequences of maximal supply limitations.

The study of metabolic fuel provision and its regulation has reached an exciting stage where specific molecular events can be correlated with parameters of the organism's ecology. This paper examines substrate supply pathways from storage sites to locomotory muscle mitochondria and discusses ecological implications of the limits for maximal flux through these pathways. The relative importance of the different oxidative fuels is shown to depend on aerobic capacity. Very aerobic, endurance-adapted animals such as long distance migrants favor the use of lipids and intramuscular fuels over carbohydrates and circulatory fuels. The hypothesis of functional co-adaptation between oxygen and metabolic fuel supply systems allows us to predict that the capacity of several biochemical processes should be scaled with maximal oxygen consumption. Key enzymes, transmembrane transporter proteins, glucose precursor supply and soluble fatty acid transport proteins must all be geared to support higher maximal glucose and fatty acid fluxes in aerobic than in sedentary species.

Aerobiosis↗
PubMed1992

Primary structure of the murine adenovirus type 1 proteinase.

The DNA sequence of an open reading frame (ORF) corresponding to the murine adenovirus type 1 (Mav1) proteinase gene was determined. 1162 base pairs were sequenced from the downstream end of the SmaI-D Mav1 genomic fragment. The sequence defines the 204 amino acid proteinase, which apparently does not possess the usual L3 polyadenylation signal, but instead the sequence AAATAA. This gene is followed by a 147 amino acid C-terminal portion of the DNA-binding protein, encoded by the complementary strand.

Adenoviridae↗
PubMed1992

Factors affecting homologous overexpression of the Saccharomyces cerevisiae lanosterol 14 alpha-demethylase gene.

The Saccharomyces cerevisiae Lanosterol 14 alpha-demethylase (14DM) gene was overexpressed in S. cerevisiae using promoter sequences of the highly expressed S. cerevisiae glyceraldehyde-3-phosphate dehydrogenase (TDH3) gene. To investigate factors affecting 14DM overproduction, the levels of 14DM-specific RNAs, apoprotein, and heme protein, respectively, were determined and the 14DM-specific RNA levels compared with the RNA levels originating from the endogenous TDH gene(s). The quantitative measurements revealed that the 14DM steady-state RNA levels reached were some three- to five-fold below the theoretically expected values. With a view towards further improving expression of the 14DM gene, the spacing between the TDH3 promoter and the AUG was adjusted precisely and to rule out possible toxic effects exerted by the 14DM protein, the TDH3 promoter was placed under galactose regulation by introducing an UASG segment. Furthermore, the effects of the gene copy number on 14DM overproduction were investigated. From the analysis of the improved expression constructs five conclusions could be reached: (1) expression from the native 14DM gene is comparable to the expression driven by the TDH3 promoter-14DM fusion construct on single copy plasmid vectors; (2) expression from the TDH3 promoter-14DM construct on single-copy vectors is nearly as efficient as expression from the corresponding endogenous TDH3 gene; (3) the gene copy number has an effect on the relative expression levels of the TDH3 promoter-14DM constructs; (4) the steady-state amounts of protein produced are very nearly proportional to gene dosage; and (5) protein toxicity does not have a major impact on 14DM production. The maximum yield of 14DM was in the order of 7% of the total yeast protein and the maximum production of functional 14DM heme protein appears to be limited by the availability of heme.

Base Sequence↗
PubMed1992

Nucleotide and deduced amino acid sequence of the canine adenovirus type 1 proteinase.

The DNA sequence of an open reading frame (ORF) corresponding to the canine adenovirus type 1 (Can1) proteinase gene was determined. A total of 1171 base pairs were sequenced from the downstream end of the HindIII-A Can1 genomic fragment, including adjacent regions corresponding to the carboxy-terminal portions of the hexon and the DNA-binding proteins. The predicted Can1 proteinase consists of 206 residues (23,325 D) of which 68% are identical and 83% are similar to the sequence of the human Ad2 proteinase. Alignment with the Ad2 proteinase identified a number of conserved residues that could form part of the catalytic triad of the enzyme.

Adenoviridae↗
PubMed1992

Adenovirus transformation revertant resistant to retransformation by E1 but not by SV40-T and HPV16-E7 oncogenes.

We have previously described a revertant cell line which expresses a dominant tumor suppressor phenotype to E1 but not to heterologous oncogenes such as c-myc, N-ras, or polyoma middle t (Sircar et al. (1988) Oncogene 3, 725-728). DNA tumor virus oncogenes have been suggested to transform cells via the common mechanism of sequestering the Rb-105 antioncoprotein. This paradigm would predict that our revertant cell line, which is resistant to retransformation by E1a, should also be resistant to the other members of the Rb-105 binding family of oncogenes. To test this hypothesis we transfected the revertant cell line with plasmids bearing SV40-T or the HPV16-E7 oncogenes. Because transformation was obtained by both oncogenes at efficiencies similar to the transformation of a related revertant cell line, the results suggest that the resistance phenotype is specific to E1a. This specificity was further confirmed by cell fusion experiments.

Adenoviridae↗
PubMed1992

Reduction of c-myc expression correlated with E1a expression but not with the transformed phenotype.

The adenovirus E1a oncogene has both positive and negative regulatory effects on the expression of a variety of host genes. Both type of effects have been reported for certain cell cycle genes such as c-myc. To study the potential role of c-myc in adenovirus transformation, we have assessed the steady-state levels of c-myc mRNA after serum stimulation in genetically related transformed or non-transformed cell lines in the presence or absence of E1a. Serum stimulated the accumulation of stable c-myc mRNA only in cell lines which did not express E1a. Therefore under the present assay conditions, E1a had a negative effect on the steady-state level of c-myc mRNA. Surprisingly, this effect was independent of the transformed phenotype.

Adenoviridae↗
PubMed1991

An erythromycin derivative produced by targeted gene disruption in Saccharopolyspora erythraea.

Derivatives of erythromycin with modifications at their C-6 position are generally sought for their increased stability at acid pH, which in turn may confer improved pharmacological properties. A recombinant mutant of the erythromycin-producing bacterium, Saccharopolyspora erythraea, produced an erythromycin derivative, 6-deoxyerythromycin A, that could not be obtained readily by chemical synthesis. This product resulted from targeted disruption of the gene, designated eryF (systematic nomenclature, CYP107), that apparently codes for the cytochrome P450, 6-deoxyerythronolide B (DEB) hydroxylase, which converts DEB to erythronolide B (EB). Enzymes normally acting on EB can process the alternative substrate DEB to form the biologically active erythromycin derivative lacking the C-6 hydroxyl group.

Amino Acid Sequence↗
PubMed1991

Complementation of adenovirus early region 1a and 2a mutants by Epstein-Barr virus immortalized lymphoblastoid cell lines.

Human B-lymphocytes may be infected by both adenoviruses and the Epstein-Barr virus (EBV). Some of the immediate early and early proteins in the two viruses are similar in function even though their primary structures are different. As these viruses might infect the same B-cells in man, we asked if complementation could take place. The adenovirus mutant H5ts125 has a thermolabile DNA-binding protein and is defective in DNA replication at 39 degrees. Several EBV-transformed human lymphoblastoid cell lines and a tamarin cell line B95-8 were infected with H5ts125 and incubated at either the nonpermissive or the permissive temperatures. Adenoviral DNA replication and assembly of new virions were observed at both temperatures, suggesting complementation by the resident EBV gene products. The adenovirus E1a region is deleted in the mutant d1312. Complementation of this mutant was only obtained in the EBV producer B95-8 cells. Immortalization by EBV was apparently not sufficient for effective complementation. This supports an earlier observation that one of the EBV early proteins (MS-EA) behaves like adenovirus E1a and can transactivate the E4 promoter in a CAT assay. The complementation of mutant adenoviruses in EBV-transformed lymphocytes may help the rescue of new adenovirus serotypes in immunosuppressed patients.

Adenovirus Early Proteins↗
PubMed1991

Effect of endurance swimming on the lactate kinetics of rainbow trout.

The lactate turnover rate of rainbow trout (Oncorhynchus mykiss) was measured by bolus injection of [U-14C]lactate at rest and during prolonged swimming at 85% Ucrit to determine the importance of this metabolic fuel for endurance locomotion in fish, to assess whether lactate exchange between white and red muscle could be a possible mechanism for supplying oxidizable fuel to their lateral red muscle, and to compare the contribution of lactate to total energy provision between teleost and mammalian species. Turnover rate only increased from 4.41 +/- 0.33 to 9.71 +/- 1.69 mumol kg-1 min-1 between rest and prolonged swimming, and the contribution of lactate oxidation to total metabolism declined during exercise. Lactate exchange between white and red muscle is, therefore, not a significant mechanism to fuel the active lateral red musculature during prolonged swimming. The lactate turnover rate of teleosts is one or two orders of magnitude lower than in mammals of equivalent size, but lactate has the same importance as a fuel in both vertebrate groups. However, lactate turnover rate and oxidation rate do not scale with body mass in the same fashion as does metabolic rate. The slope of the mammalian relationship for whole-body lactate turnover and oxidation is much lower (0.58) than the slope of the classic relationship for metabolic rate (0.75), indicating that lactate is a much more important oxidative substrate for small than for large animals.

Animals↗
PubMed1991

Dominant suppression of adenovirus mediated transformation and insufficiency of p105Rb binding as a condition for oncogenic transformation.

An adenovirus-specific transformation resistant cell line (G2) expressing biologically active E1a proteins and originally isolated as a revertant from Ad2-transformed rat cells (F4), was shown to form stable Rb-E1a and 300K-E1a complexes in immunoprecipitation experiments. Consistent with the transformation resistant phenotype, cell hybrids between G2 and F4 were all nontumorigenic. Retrovirus insertion mutagenesis resulted in tumorigenic cell lines and identified a common locus responsible for the E1a-specific dominant tumor suppressor phenotype of G2 cells.

Adenoviruses, Human↗
PubMed1990

Drug-induced revertants of adenovirus-transformed cells: retransformation by 5-azacytidine without reactivation of E1a.

We have isolated drug-resistant variants from adenovirus-transformed rat cells that had concomitantly lost their transformed phenotype. Our aim was to determine the reason for reversion, to attempt retransformation with 5-azacytidine (5-AzaC) and to study the mechanism of retransformation. Of the three cell lines studied, one (G4F) had lost the integrated E1a genes, whereas the other two (G2a and G5) failed to synthesize E1a RNA or proteins. Incubation of these cell lines with 3 microM-5-AzaC for 2 days, followed by passaging in the absence of drug, gave rise to transformed foci in all of the cell lines. The efficiency of transformation was typical of each cell line. Surprisingly, retransformation was not accompanied by the reappearance of detectable levels of E1a gene activity in the G2aAza and G5Aza cell lines. In search of a mechanistic explanation for the loss of gene activity in the revertants and its reappearance in the retransformants, we examined the state of methylation of the E1a gene region in these cells. Neither the E1a promoter nor its upstream region was methylated in the revertants or the 5-AzaC retransformants. These results suggest that E1a transcription was suppressed by mechanisms other than DNA methylation and that 5-AzaC could retransform these cells without lifting the E1a-suppressed state.

Adenovirus Early Proteins↗
PubMed1990

Role of triglyceride-fatty acid cycle in controlling fat metabolism in humans during and after exercise.

We have investigated the role of triglyceride-fatty acid cycling in amplifying control of the net flux of fatty acids in response to exercise and in recovery from exercise. Five normal volunteers were infused with [1-13C]palmitate and D-5-glycerol throughout rest, 4 h of treadmill exercise at 40% maximum O2 consumption, and 2 h of recovery. Total fat oxidation was quantified by indirect calorimetry. Lipolysis (rate of appearance of glycerol) increased from 2.1 +/- 0.3 to 6.0 +/- 1.2 mumol.kg-1.min-1 after 30 min of exercise and progressively increased thereafter to a value of 10.5 +/- 0.8 mumol.kg-1.min-1 after 4 h. Lipolysis decreased rapidly during the first 20 min of recovery, but it was still significantly elevated after 2 h of recovery. The rate of appearance of free fatty acids followed the same pattern of response. Seventy percent of released fatty acids were reesterified at rest, and this value decreased to 25% within the first 30 min of exercise. Reesterification remained less than 35% of lipolysis until the start of recovery, at which time the value rose to 90%. In exercise, more than one-half the increase in fat oxidation could be attributed to the reduction in the percent reesterification. Most of the change in percent reesterification during exercise and recovery was caused by changes in extracellular cycling of fatty acids released into plasma. We conclude that triglyceride-fatty acid cycling plays an important role in enabling a rapid response of fatty acid metabolism to major changes in energy metabolism.

Adult↗
PubMed1976

Factors influencing the production of interferon from L cells.

The production of interferon from Newcastle disease virus-infected mouse L929 cells was investigated in relation to superinduction procedures, cell density, cellular cyclic adenosine 3',5'-monophosphate (cAMP) levels, and rate of incorporation of 14C-labeled protein hydrolysate into trichloroacetic acid-precipitable material. Densely populated cultures did not have their interferon production enhanced through "superinduction" using cycloheximide, actinomycin D, or the two antimetabolites in combination. These dense cultures produced more interferon per cell than less dense cultures, even though the interferon production from the latter cells could be enhanced two- to threefold by cycloheximide or combined cycloheximide and actinomycin D. Cells in densely populated cultures relative to those from sparsely populated cultures were smaller in volume, had a correspondingly reduced protein content and a lower concentration of cAMP, and were less able to concentrate 14C-labeled protein hydrolysate, although proportionally they were just as efficient in incorporating labeled precursors into trichloroacetic acid-precipitable polypeptides.

Animals↗