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Y Edwards

Publications and source records attributed to Y Edwards.

29 records · Page 2Linked to original sources
PubMed1988

Changes in skeletal muscle gene transcription induced by chronic stimulation.

The mRNA products of four genes, carbonic anhydrase III (CAIII), fast myosin heavy chain (MHCf), actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), were assayed by Northern and slot-blot analysis in rabbit tibialis anterior and extensor digitorum longus muscles that were subjected to electrical stimulation for periods of up to 21 days. Marked changes in mRNA levels were seen for all four genes. The mRNA for CAIII, which is specific for type 1 fibers, rose significantly, whereas the MHCf mRNA fell markedly in the stimulated muscles. Changes in GAPDH mRNA were consistent with a reduced dependence on anaerobic glycolysis as an energy source. Actin mRNA levels were noticeably depressed in the early stages of stimulation. Thus for several classes of muscle protein, the response to chronic low-frequency stimulation appears to involve changes at the level of gene transcription.

Actins↗
PubMed1987

Deletions of fetal and adult muscle cDNA in Duchenne and Becker muscular dystrophy patients.

We have isolated a cDNA molecule from a human adult muscle cDNA library which is deleted in several Duchenne muscular dystrophy patients. Patient deletions have been used to map the exons across the Xp21 region of the short arm of the X chromosome. We demonstrate that a very mildly affected 61 year old patient is deleted for at least nine exons of the adult cDNA. We find no evidence for differential exon usage between adult and fetal muscle in this region of the gene. There must therefore be less essential domains of the protein structure which can be removed without complete loss of function. The sequence of 2.0 kb of the adult cDNA shows no homology to any previously described protein listed in the data banks although sequence comparison at the amino acid level suggests that the protein has a structure not dissimilar to rod structures of cytoskeletal proteins such as lamin and myosin. There are single nucleotide differences in the DNA sequence between the adult and fetal cDNAs which result in amino acid changes but none that would be predicted to change the structure of the protein dramatically.

Amino Acid Sequence↗
PubMed1987

Isolation of a cDNA probe for a human jejunal brush-border hydrolase, sucrase-isomaltase, and assignment of the gene locus to chromosome 3.

We report the nucleotide sequence and derived amino acid sequence of a cDNA clone encoding most of the N-terminal, isomaltase region of human sucrase-isomaltase (SI). A plasmid containing this cDNA, pS12, identifies a 6-kb mRNA found in human jejunum and the human colon carcinoma cell line Caco-2. This human SI cDNA shows extensive overall homology with recently published rabbit SI cDNA. Using pS12 to probe DNA from a panel of somatic cell hybrids, we have assigned the gene encoding human SI to chromosome 3.

Amino Acid Sequence↗
PubMed1986

Multiple mRNAs for human alcohol dehydrogenase (ADH): developmental and tissue specific differences.

Human class I alcohol dehydrogenase (ADH) genes show developmental and tissue specific differences in expression at the polypeptide level. In these studies ADH expression was investigated at the RNA level. Northern blot analysis of total and poly (A) RNA from adult liver using pADH12 probe demonstrated multiple RNA size classes of 2.6, 2.2, 1.9 and 1.6kb. In contrast, fetal liver, and fetal intestine contained only 2.6 and 1.6kb mRNA while fetal lung showed only 2.6kb mRNA. All of these tissues showed a relative reduction in the amount of ADH mRNA present when compared to adult liver. Immunoprecipitation of in vitro translation products of adult liver RNA by polyclonal ADH antibody revealed a single polypeptide of 40,000 daltons. This result points out the homogeneity of size of class I ADH polypeptides despite mRNA size diversity. Variation in length of the 3' untranslated region probably contributes to the multiple size classes of ADH mRNA observed.

Alcohol Dehydrogenase↗
PubMed1985

The expression of human glycerol-3-phosphate dehydrogenase in human/rodent somatic-cell hybrids.

Our previous studies using rodent/human somatic-cell hybrids suggested that the expression of human mitochondrial glycerol-3-phosphate dehydrogenase (GPDM) is dependent on the presence of human mitochondria. This has now been tested directly by analysis of GPDM activity in a series of nine hybrid-cell lines, four segregating human chromosomes and five losing rodent chromosomes (reverse segregants). The chromosome composition of the hybrids was deduced from analysis of biochemical markers and examination of G- and G11-banded metaphase spreads and the mitochondrial content was determined by Southern blot analysis, using cloned mouse and human mtDNA sequences as probes. We found that the mtDNA species present in these hybrids correlated exactly with the pattern of chromosome segregation such that the conventional hybrids contained rodent mtDNA and the reverse segregants human mtDNA. However, the pattern of GPDM expression was not directly correlated with the species of chromosomes or mitochondria present: all the hybrids showed strong rodent GPDM activity and two from each class of hybrid also showed human GPDM activity but the other hybrids were negative for human GPDM. We conclude that rodent GPDM readily integrates into human mitochondria, that the expression of rodent GPDM is not dependent on the presence of rodent mitochondria, and that GPDM is not coded by mtDNA. Human GPDM either is not capable of being inserted into the rodent mitochondrial membrane or is regulated in some way in the hybrid cells by an unidentified rodent factor.

Animals↗
PubMed1984

The 3'-untranslated sequence of human skeletal muscle alpha-actin mRNA.

A human actin cDNA clone pGF3 isolated from a fetal skeletal muscle cDNA library is described. The insert cDNA is homologous to skeletal muscle alpha-actin as judged by restriction mapping and nucleotide sequencing. The recombinant contains a substantial portion of the coding and the complete 3'-untranslated region. Comparison of the 3' ends of human and rat skeletal muscle and human cardiac alpha-actins reveals little homology between different types of actin genes in man but marked conservation of this region in the skeletal muscle actins of man and rat.

Actins↗
PubMed1981

Human succinate dehydrogenase: biochemical and genetic characterization.

A simple procedure for the preparation of soluble human succinate dehydrogenase is described. These preparations have proved suitable for analysis by zone electrophoresis, using a specific stain to detect activity after separation. In a survey of succinate dehydrogenase from various tissues and different individuals, no evidence for genetic heterogeneity due to the expression of either multiple loci or alternative alleles at the succinate dehydrogenase locus was found. However, epigenetic heterogeneity in both molecular size and charge was seen and various explanations for the occurrence of the isoenzymes are explored. Estimates of molecular size (93,300 +/- 9100) suggest that the smallest active unit of succinate dehydrogenase accounts for the major part of the solubilized activity. Kinetic studies have shown that the apparent Km values for succinate (0.9 mM) and PMS (0.4 mM) are comparable to those previously described for the beef heart enzyme, and these parameters were not significantly altered when the enzyme was removed from the membrane milieu. However a marked non-succinate-dependent activation of the membrane-associated enzyme at 38 C is apparently lost on solubilization, and this observation may have some bearing on earlier reports of an apparent decrease in Vmax on solubilization of succinate dehydrogenase.

Adult↗
PubMed1980

Distribution of CAIII in fetal and adult human tissue.

Carbonic anhydrase III (CAIII), an enzyme recently shown by conventional electrophoresis to be muscle specific, has been qualified by "rocket" immunoelectrophoresis. This more sensitive technique has shown that the enzyme is virtually specific to skeletal muscle, where it occurs at a level of 5 mg per g, with trace levels in smooth muscle, cardiac muscle, and lung. In man there does not appear to be any correlation between CAIII levels and the proportion of red and white muscle fibers. The fetal development of CAIII has also been examined using immunoelectrophoresis, and the enzyme can be detected at 11 weeks' gestation. The CAIII level rises gradually up to 25 weeks, and there is then a more dramatic increase to reach approximately half adult level at birth.

Adult↗
PubMed1979

Characterization of human carbonic anhydrase III from skeletal muscle.

A third form of human carbonic anhydrase (CA III), found at high concentrations in skeletal muscle, has been purified and characterized. This isozyme shows relatively poor hydratase and esterase activities compared to the red cell isozymes, CA I and CA II, but is similar to these isozymes in subunit structure (monomer) and molecular size (28,000). CA III is liable to posttranslational modification by thiol group interaction. Monomeric secondary isozymes, sensitive to beta-mercaptoethanol, are found in both crude and purified material and can be generated in vitro by the addition of thiol reagents. Active dimeric isozymes, generated apparently by the formation of intermolecular disulfide bridges, also occur but account for only a small proportion of the total protein and appear only when the concentration of CA III is particularly high.

Adult↗