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J Boulter

Publications and source records attributed to J Boulter.

49 records · Page 3Linked to original sources
PubMed1985

alpha-Fetoprotein and albumin gene expression in brain and other tissues of fetal and adult rats.

Quantitative measurement of messenger RNA (mRNA) for alpha-fetoprotein (AFP) and albumin in developing rat liver and in different fetal and adult tissues reveals a close correlation between the previously determined rate of protein synthesis and mRNA levels. mRNA for AFP and albumin exists in fetal intestine, lung, liver and kidney whereas there are no such transcripts in fetal brain or heart. There are no mRNA transcripts for AFP in any adult organs other than the liver. The lack of mRNA AFP in fetal brain tissue indicates that the AFP found in fetal brain cells is absorbed from the serum. This finding supports the hypothesis that AFP may serve as a carrier protein to deliver bound molecules, such as non-esterified fatty acids, to brain cells at a specific time during development.

Albumins↗
PubMed1985

Lack of correlation of methylation and alphafetoprotein and albumin gene expression during liver growth, in hepatocellular carcinomas, and during hepatocarcinogenesis.

Analysis of AFP and albumin genes fails to demonstrate a correlation of gene activity with the degree of gene methylation as determined by restriction endonuclease fragments obtained using the isoschizomer pair, Hpa II and Msp I. There is no difference in the methylation patterns of DNA from high and low producing tissues, such as fetal and adult liver for AFP, hepatoma 7777 and 9098 for AFP; adult lung, brain, kidney and liver for albumin; fetal liver and brain or heart for AFP, etc. Minor selective differences in gene methylation that correlate with AFP or albumin gene expression cannot be ruled out.

Albumins↗
PubMed1985

Isolation of a clone coding for the alpha-subunit of a mouse acetylcholine receptor.

The mouse cell line BC3H-I synthesizes an acetylcholine receptor (AChR) with the pharmacological properties of a muscle nicotinic cholinergic receptor. We have purified mRNA from this cell line and used the size-fractionated poly(A)+RNA to produce a cDNA library of approximately 50,000 clones. The library was screened with a subclone containing genomic sequences coding for the putative acetylcholine-binding site of the alpha-subunit of chicken AChR. We obtained a plasmid, pMAR alpha 15, with a 1,717-base pair insert. The insert cDNA has 26 nucleotides at the 5'-end which code for a portion of the signal peptide followed by a single open reading frame of 1,311 nucleotides which code for a protein of 49,896 daltons. The insert has 377 bases of 3'-untranslated sequence with 3 polyadenylation sites. Radiolabeled plasmid DNA has been used to identify homologous RNA species of about 2 kilobases in Northern blot analyses of poly(A)+ selected RNA from BC3H-I cells. A similar size mRNA is seen in innervated mouse diaphragm and leg muscle, and both mouse and rat brain. Comparisons of the deduced amino acid sequence of the mouse AChR alpha-subunit with Torpedo marmorata, T. californica, chicken, human, and calf sequences show overall homologies of 80%, 80%, 86%, 96%, and 95%, respectively. More detailed analyses reveal a non-random distribution of amino acid substitutions in several structural domains. Based on the absolute conservation of cysteine residues, a new model for the arrangement of the disulfide bonds in the extracellular portion of the alpha-subunit is proposed.

Amino Acid Sequence↗
PubMed1985

Muscle denervation increases the levels of two mRNAs coding for the acetylcholine receptor alpha-subunit.

The mRNA coding for the alpha-subunit of the acetylcholine receptor was studied in mouse leg and rat diaphragm muscle. We find that denervation of rat diaphragm results in a 7-fold increase in mRNA coding for the alpha-subunit, whereas denervation of mouse leg muscle results in approximately a 50-fold increase in alpha-subunit-specific mRNA. The relationship of the mRNAs purified from innervated and denervated muscle was investigated by SI nuclease mapping. Two mRNA species were found in both innervated and denervated muscle which differ in their 3'-untranslated region. The levels of both these mRNA species increase upon denervation of mouse leg muscle.

Animals↗
PubMed1984

Polymorphism of rat alphafetoprotein and albumin genes.

Restriction endonuclease digestion using Hind III and Msp I and Southern blot analysis of DNA from the liver of three inbred rat strains and one outbred strain using cDNA probes yields two banding patterns for the alphafetoprotein and albumin genes. Buffalo and Fischer DNA have one pattern whereas ACI had a different pattern for both genes. Sprague Dawley DNA contains fragments of both patterns suggesting heterozygosity in some individuals of this strain. These polymorphisms do not appear to be associated with any structural or biological differences in the proteins resulting from expression of these genes.

Albumins↗
PubMed1979

Concanavalin A inhibition of alpha-bungarotoxin binding to a nonfusing muscle cell line.

Incubation of a nonfusing muscle cell line, BC3H1, with concanavalin A (Con A) results in a maximum decrease of 35% in the cell's ability to bind alpha-bungarotoxin (alpha-BuTx). The Con A-induced inhibition of 125I-alpha-BuTx binding is reversible and the degree of inhibition parallels the degree of saturation of Con A binding sites on the cell surface. The maximum level of Con A-induced inhibition of 125I-alpha-BuTx binding is not affected by increasing the time of incubation in Con A, using higher concentrations of Con A or by increasing the time of incubation in the presence of 125I-alpha-BuTx. In addition, all BC3H1 cells in culture are sensitive to the Con A-induced inhibition of 125I-alpha-BuTx binding. A comparison of the pseudo-first order rate constants for 125I-alpha-BuTx binding to untreated (8.6 x 10(4) M-1 S-1) and Con A-treated (5.4 x 10(4) M-1 S-1) BC3H1 cells, however, shows that those acetylcholine receptors in Con A-treated cells which bind 125I-alpha-BuTx do so with a lowered apparent affinity. Partial inhibition of toxin-binding capacity is not a consequence of two classes of acetylcholine receptors on the cell surface. Furthermore, individual receptors experience partial inhibition of their binding capacity by Con A, resulting in receptors with at least one binding site blocked and at least one site available for alpha-BuTx binding.

Acetylcholine↗
PubMed1975

Isolation of specialized transducing bacteriophage lambda carrying genes of the L-arabinose operon of Escherichia coli B/r.

A heat-inducible lysis-defective phage lambda (lambdacI857S7) has been integrated at multiple sites within the L-arabinose region (araCOIBAD) of a strain of Escherichia coli K-12 deleted for the normal lambda attachment site (lambdaattdelta). The lambda phage has become integrated with opposite orientations at two different loci within the aratb gene and with the "normal" orientation (clockwise N-RA-J) at a single site in the araC gene. The burst size, spontaneous-curing frequencies, and number of prophage harbored by each of the ara secondary-site lysogens have been determined. From these secondary-site lysogens it has been possible to generate plaque-forming ara-transducing phage (lambdapara) and defective ara-transducing phage (lambdadara), as well as defective leucine-transducing particles (lambdadleu). The construction and characterization of these lambdaara-transducing phage and their derivatives which carry genetically defined portions of the L-arabinose region are presented.

Arabinose↗
PubMed1974

Direction of transcription of the regulatory gene araC in Escherichia coli B-r.

The protein product of the regulatory gene araC can be synthesized in a cell-free, protein-synthesizing system programmed with a lambdaparaC(+)B DNA template. Hybrid, renatured phage DNA molecules prepared with DNA from phages lambdaparaC(+)B and lambdaparaC3B (araC3 is a nonsense mutation) were used to program the cell-free synthesis of the araC protein. The findings observed lead to the conclusion that the codogenic strand of the araC gene is on the light strand of the phage DNA. The araB gene is on the heavy strand, as determined by DNA.RNA hybridization. Thus, with regard to the standard E. coli map, araC is transcribed in a clockwise direction, whereas transcription of the araBAD operon has a counterclockwise orientation. The technique described should allow one to determine the direction of transcription of any gene that can be incorporated into the genome of a specialized transducing phage.

Arabinose↗
PubMed1974

Metabolism of D-arabinose by Escherichia coli B-r.

The pathway for d-arabinose metabolism in Escherichia coli B/r has been determined. Evidence is presented to support the following metabolic scheme: d-arabinose d-ribulose right harpoon-up d-ribulose-5-phosphate d-xylulose-5-phosphate.

Aldehyde-Lyases↗
PubMedDate not supplied

Isolation of a cDNA clone coding for a possible neural nicotinic acetylcholine receptor alpha-subunit.

We have isolated a complementary DNA clone containing sequences homologous to those encoding the alpha-subunit of a mouse muscle nicotinic acetylcholine receptor. Based on the structural similarities between the encoded protein and the muscle acetylcholine receptor alpha-subunit, and the presence of hybridizing RNA species in the brain, we propose that this clone codes for a neural nicotinic acetylcholine receptor alpha-subunit.

Amino Acid Sequence↗