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J T Epplen

Publications and source records attributed to J T Epplen.

At least 325 records · Page 18Linked to original sources
PubMed1983

The primitive code and repeats of base oligomers as the primordial protein-encoding sequence.

Even if the prebiotic self-replication of nucleic acids and the subsequent emergence of primitive, enzyme-independent tRNAs are accepted as plausible, the origin of life by spontaneous generation still appears improbable. This is because the just-emerged primitive translational machinery had to cope with base sequences that were not preselected for their coding potentials. Particularly if the primitive mitochondria-like code with four chain-terminating base triplets preceded the universal code, the translation of long, randomly generated, base sequences at this critical stage would have merely resulted in the production of short oligopeptides instead of long polypeptide chains. We present the base sequence of a mouse transcript containing tetranucleotide repeats conserved during evolution. Even if translated in accordance with the primitive mitochondria-like code, this transcript in its three reading frames can yield 245-, 246-, and 251-residue-long tetrapeptidic periodical polypeptides that are already acquiring longer periodicities. We contend that the first set of base sequences translated at the beginning of life were such oligonucleotide repeats. By quickly acquiring longer periodicities, their products must have soon gained characteristic secondary structures--alpha-helical or beta-sheet or both.

Amino Acid Sequence↗
PubMed1982

Base sequence of a cloned snake W-chromosome DNA fragment and identification of a male-specific putative mRNA in the mouse.

A 2.5-kilobase fragment of a sex-specific satellite DNA from the Colubrid snake species Elaphe radiata has been cloned, and its sequence has been determined. It contains 26 and 12 copies, respectively, of two base quadruplets, G-A-T-A and G-A-C-A, as its sole highly repetitious elements. Southern hybridization experiments with genomic DNA of the chicken, the mouse, and man indicated male sex-specific conservation of at least parts of this cloned DNA. In situ hybridization experiments with metaphase chromosomes of the mouse showed that elements that can cross-hybridize with parts of the cloned snake DNA are concentrated in the pericentric region of the Y chromosome. In blot hybridization experiments with liver poly(A)+ polysomal RNAs of male and female mice, a probe consisting of the first 1,224 bases of the cloned snake DNA singled out a male-specific RNA of 1,250-1,400 bases. Inasmuch as the proximal end of this probe contained an open reading frame (44 consecutive amino acid-specifying codons), the male-specific putative mRNA so detected may specify H-Y antigen. By contrast, a probe consisting of bases 1,480-1,906, containing the simple repeats of the quadruplets, singled out a shorter (approximately 1,000-base) RNA from males and females alike. Although this RNA is poly(A)+, we have yet to establish its attachment to ribosomes.

Animals↗
PubMed1982

Identification of the 45-base-long primordial building block of the entire class I major histocompatibility complex antigen gene.

Within the 885-base-long incomplete coding sequence for the mouse class I major histocompatibility complex (MHC) antigen H-2Kb heavy chain, we have identified 32 recurring base oligomers (1 decamer, 4 octamers, 9 heptamers, and 18 hexamers), the decamer being T-C-A-C-C-C-T-G-A-G. The compilation of these recurring base oligomers led to the identification of the 45-base-long primordial building block for this class of genes which is: C-C-T-G-C-G-G-A-C-A-A-T-G-T-C-A-C-C-C-T-G-A-G-C-T-G-C-T-G-G-G-C-T-C-T-G-G-G-C-G - T-T-G-A-C. Inasmuch as these base oligomers that recurred within the mouse H-2Kb coding sequence proper also recurred within the adjacent, transcribed noncoding sequence, we conclude that the entire approximately equal to 4,000-base-long germ-line DNA segment containing eight or nine separate exons of the coding sequence for class I MHC antigen heavy chains can be construed as having evolved from tandem repeats of this one 45-base-long primordial building block.

Amino Acid Sequence↗
PubMed1982

Patterns of cyclic AMP phosphodiesterases in the rat pineal gland: sex differences in diurnal rhythmicity.

Rat pineal cyclic AMP (cAMP) phosphodiesterases (PDEs) were studied during male development and in adult females by means of gel electrophoresis, kinetic analysis, and diurnal activity measurements. In both male and female rats, two molecular forms of PDE coexisted. In neonatal-infantile males (birth to day 20 of age) a third isozyme was detected. The kinetic properties of PDEs were essentially similar in infantile and adult male rats with respect to the Kms; however, a minimal Vmax was observed at 10.00 a.m. in adults, but not in neonatal animals. PDE activity in male rats was maximal between 2.00 a.m. and 4.00 a.m., and also at 4.00 p.m., and was minimal at 10.00 a.m. This pattern appeared to develop during the infantile period of life. The pattern of activity observed in adult female rats was similar to that of adult males, except that it lacked a pronounced afternoon peak. The diurnal PDE activity profile of castrated male adult rats was similar to female adults and testosterone substitution in male castrated rats resulted in a pattern which resembled that of adult male controls. In addition to the above measurements, testosterone serum levels have been determined during all of these experiments.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1981

Cyclic AMP phosphodiesterase in Leydig cell preparations of the rat testis.

Enzymatic digestion of the interstitial tissue of early juvenile and adult rat testes resulted in an enrichment of the Leydig cell population. The cells of the intertubular preparation from adult testes were separated by centrifugal elutriation, according to differences in sedimentation velocity, a counter-flow centrifugation technique leading to 70% Leydig cell purity. Using this approach, it was possible to demonstrate that Leydig cells from adult testes contain only low affinity isoenzymes of cyclic AMP phosphodiesterase (PDE; E.C.: 3.1.4.17), an intracellular regulator of cAMP. Starch gel electrophoresis showed that the isozyme of cAMP PDe of Leydig cells is masked in crude testis homogenates due to the relatively low level of these cells in the total population. In Leydig cells, there are two different electrophoretic forms expressed which resemble two of eleven different molecular forms of cAMP PDE demonstrated for comparison in 21 different organs of the adult rat. An interstitial cell preparation from early juvenile testes, with a Leydig cell content of up to 20%, was also investigated electrophoretically with regard to molecular forms of cAMP PDE, the properties of which were characterized by kinetics analysis of cAMP hydrolysis. The results presented are discussed in relation to the onset of testosterone synthesis in Leydig cells of prepubertal rats leading to the initiation of male puberty.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1980

Patterns of cyclic AMP phosphodiesterases during rat testis development.

Cyclic AMP phosphodiesterase (PDE; E.C.: 3.1.4.17) plays a crucial role in the regulation of intracellular cAMP levels arising from the hormonal activation of membrane-bound adenylcyclase in the target cell. In the present study, we revealed a complex sequence of appearance and disappearance of individual molecular electrophoretic forms of PDE during the development of the rat testis. Kinetic analysis of cAMP hydrolysis in crude testis homogenates of developmental stages where multiple PDE isozymes are expressed revealed complex kinetic behavior of PDE. After separation of individual isozymes by elution from starch gel blocks, several enzymatic forms still act with complex kinetics, indicating negatively cooperative behavior. The stage specificity of the kinetic properties of PDE appears to be related to the hormonally regulated events leading to the initiation of male puberty.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1980

Sequence organization of animal nuclear DNA.

Animal nuclear genomes contain DNA sequences of various degrees of repetition. These sequences are organized in highly ordered fashions; repetitive and nonrepetitive sequences either alternate in short periods, i.e., short [0.2-0.4 kilobases (kb) long] repeats are flanked by nonrepetitive sequences less than 2 kb long, or in longer periods, with repetitive and/or nonrepetitive sequences extending for several kilobases. There are two main categories of genome organization, namely those exhibiting short-period interspersion and those that do not. There are arguments for and against a regulatory role of short interspersed repetitive sequences. Besides the merely 'statistical' kinetic approach by conventional reassociation kinetics, sequence organization has been studied by restriction endonuclease mapping and nucleotide sequencing. Such studies have revealed some general features of the organization of the eukaryotic gene and its transcripts, namely possible 'promoters', 'leaders', 'introns', 'exons', 'flanking sequences', 'caps', ribosome-binding sites, and poly(A) sequences. This paper discusses how these elements of a gene might serve regulatory roles in its expression.

Animals↗
PubMed1980

Cyclic AMP phosphodiesterases of the rat ovary. Oestrous cycle dependent activity change of high affinity form.

In the rat ovary, from prenatal to adult stages, two electrophoretically distinct forms of cyclic AMP phosphodiesterase (E.C.: 3.1.4.17) occur. These forms were analysed for their affinity towards cAMP in both crude and partially purified preparations. Whereas the slow electrophoretic form, which has a low affinity towards cAMP, shows a linear Lineweaver-Burk plot, the fast electromorph exhibits anomalous kinetic behaviour in that cAMP affinity appears to increase with decreasing cAMP-concentration. This form shows dramatic alteration of maximum velocity during the oestrous cycle, attaining maximum activity at metoestrus. This rise of activity is sensitive to inhibition of protein synthesis.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1979

Contrasting DNA sequence organisation patterns in sauropsidian genomes.

The genomic DNA organisation patterns of four sauropsidian species, namely Python reticularis, Caiman crocodilus, Terrapene carolina triungius and Columba livia domestica were investigated by reassociation of short and long DNA fragments, by hyperchromicity measurements of reannealed fragments and by length estimations of S1-nuclease resistant repetitive duplexes. While the genomic DNA of the three reptilian species shows a short period interspersion pattern, the genome of the avian species is organised in a long period interspersion pattern apparently typical for birds. These findings are discussed in view of the close phylogenetic relationships of birds and reptiles, and also with regard to a possible relationship between the extent of sequence interspersion and genome size.

Alligators and Crocodiles↗
PubMed1979

Genome analysis of Amphioxus and speculation as to the origin of contrasting vertebrate genome organization patterns.

1. The genome of Amphioxus was investigated by DNA reassociation techniques for the amount of repetitive and non-repetitive sequences and its pattern of organization. 2. A comparison of the amount of non-repetitive DNA between Amphioxus and the tunicate Ciona intestinalis does not support the hypothesis that the Cephalochordates have arisen from the Tunicates by polyploidy. 3. In the Amphioxus genome repetitive and non-repetitive elements are predominantly arranged in a short period interspersion pattern. Conclusions are presented as to the evolution of contrasting genome organization patterns among vertebrates.

Animals↗
PubMed1978

DNA sequence organisation in avian genomes.

By means of renaturation kinetics of DNA of the three avian species Cairina domestica, Gallus domesticus and Columba livia domestica the following major DNA repetition classes were observed: a very fast reannealing fraction comprising about 15% of the DNA, a fast or intermediate reannealing fraction that makes up 10%, and a slow reannealing fraction of about 70%, which apparently renatures with single copy properties. --Comparing the reassociation behaviour of short (0.3 kb) and long (greater than 2 kb) DNA fragments of duck and chicken it becomes apparent that only 12% (duck) and 28% (chicken) of the single copy DNA are interspersed with repetitive elements on 2 to 3 kb long fragments. The lengths of the repetitive sequences were estimated by optical hyperchromicity measurements, by agarose A-50 chromatography of S1 nuclease resistant duplexes and by electron microscopic measurements of the S1 nuclease resistant duplexes. It was found that in the case of the chicken DNA the single copy sequences alternating with middle repetitive ones are at least 2.3 kb long; the interspersed moderate repeats have a length average of at least 1.5 kb. The sequence length of the moderate repeats in duck DNA is smaller. The results show that the duck and the chicken genomes do not follow the short period interspersion pattern of genome organisation, characteristic of the eucaryotic organisms studied so far.

Animals↗
PubMed1977

Karyotype alterations of BHK-21 cell lines by persistent virus infections I. Development of the cell lines and conventional chromosome analysis.

From BHK-21 cell cultures inoculated with different viruses cell lines with persistent infection were developed and examined for alterations in their karyotypes. In all cell lines studied, even in those no longer producing infective virus chromosomal breaks and interchanges were observed. Besides these effects probably induced directly by the virus infection, a shift in the number of chromosomes was found. Based on certain properties of the BHK-21 cell lines carrying a virus infection a hypothesis is discussed concerning the cooperation of various cellular and viral factors which enable a virus to persist.

Animals↗
PubMed1977

Karotype alterations of BHK-21 cell lines by persistent virus infections. II. Analysis of the banding patterns.

Results of banding pattern analysis of the chromosome set of cell lines derived from BHK-21 by persistent infection with different viruses (Schwöbel et al., 1977) are reported. Almost all metaphases contained several structurally altered chromosomes which could not be identified. On the other hand, for each cell line analysed a characteristic karyotype could be established. The possible relevance of the results with respect to persistent virus infections in vivo is discussed.

Animals↗