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Biomedical subjects

H Schaller

Publications and source records attributed to H Schaller.

At least 145 records · Page 8Linked to original sources

An RNA transcribed from DNA at the origin of phage fd single strand to replicative form conversion.

Phage fd DNA complexed with DNA binding protein I was used by Escherichia coli RNA polymerase (nucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) to synthesize an RNA at the origin of single strand to double strand replication. The isolated ori-RNA gave a simple fingerprint after nucleolytic digestion and has a length of about 30 nucleotides. The characterization of the oligonucleotides from the nuclease digest and the extension of the ori-RNA with DNA polymerase I and subsequent restriction of the DNA gave its exact localization in the fd genome, and its total sequence was deduced from the known DNA sequence in this region.

Bacteriophages↗

Nucleotide sequence of bacteriophage fd DNA.

The sequence of the 6,408 nucleotides of bacteriophage fd DNA has been determined. This allows to deduce the exact organisation of the filamentous phage genome and provides easy access to DNA segments of known structure and function.

Base Sequence↗

Influence of Mg2+-ions on the properties of rat heart mitochondria in dependence on the preparation.

1. The preparation of rat heart mitochondria with Potter-Elvehjem homogenizer results in mitochondria showing stimualtion of respiration induced by Mg2+. This stimualtion is neither caused by adherent hexokinase nor by energy-dependent magnesium accumulation (Mg2+ content in the presence of 10 mM glutamate: 22 nmoles/mg protein; in the presence of glutamate plus antimycin A 21 nmoles/mg protein). 2. The effect of added magnesium is excluded by addition of carboxyatractyloside. This demonstrates the activity of an ATPase outside of the mitochondrial inner membrane. 3. A simple and rapid method for the preparation of Mg2+-insensitive rat heart mitochondria is presented. The minced heart is pressed through a normal syringe and then treated with trypsin. 4. A comparison of mitochondria of both preparations shows that there is no difference in magnesium content and no energy-dependent magnesium influx.

Adenylate Kinase↗

The dependence on the extramitochondrial ATP/ADP-ratio of the oxidative phosphorylation in mitochondria isolated by a new procedure from rat skeletal muscle.

A simple method for preparation of rat skeletal muscle mitochondria is presented using gentle mechanical homogenization in a syringe and nagarse treatment (EC 3.4.4.16). This method enables the preparation of skeletal muscle mitochondria, whose outer membrane is intact to 95%. Furthermore, with mitochondria prepared by this method the regulation of respiration and phosphorylation by the extramitochondrial ATP/ADP-ratio can be demonstrated. In accordance to rat liver and heart mitochondria and to mitochondria of rabbit reticulocytes, the regulation by the extramitochondrial ATP/ADP-ratio lies in the range from 5 (corresponding to 98% of the maximum respiration) to 100 (corresponding to state 4). At extramitochondrial ATP/ADP-ratios from 0.01 to 1 the respiration rate is nearly constant (maximum rate of respiration).

Adenosine Diphosphate↗

Interaction of RNA polymerase with promoters from bacteriophage fd.

Replicative form DNA of bacteriophage fd, which had been fragmented with the restriction endonuclease II from Hemophilus parainfluenzae (endo R- HpaII), was reacted with Escherichia coli RNA polymerase; the resulting stable preinitiation complexes were analysed using the filter binding assay followed by gel electrophoresis. At 120mM KCL the first-order rate constants for complex decay were determined to be 10(-2)-10(-6)s-1. The second-order rate constants for complex formation were found to be about 10(6) -10(7) M-1 s-1. From these values association constants for the individual promoters were calculated to be 2 x 10(-8) -2 x 10(-11) M-1. The rate of formation and the stability of promoter complexes was enhanced in superhelical DNA. No evidence was found for stable promoter-specific closed complexes consisting of enzyme and helical DNA. This and the kinetic data suggest that the unwinding of base pairs is already important early in promoter selection, and not only for the formation of the final open complex. The initiation of RNA synthesis form the preinitiation complex was faster than complex dissociation and complex formation for all promoters. Consequently, the initiation efficiency of a promoter is determined by the rate of complex formation, and not by its 'affinity' for the enzyme. No correlation was found between the relative order of the fd promoters for the binding and the dissociation reaction. This is explained by different structural determinants, for the two reactions, which are located in different parts of the promoter DNA.

Base Sequence↗

[Pre-operative embolisation of vascular tumours in the head (author's transl)].

Trans-vascular, pre-operative embolisation in order to reduce blood flow was carried out with excellent results in a patient with a naso-pharyngeal fibroma, and with relatively good results in a patient with a pterion meningioma. This technique can be carried out following the diagnostic angiogram through the same catheter; it reduces operative blood loss, improves haemostasis and the surgeon's view of the operative field. The method can be used simply and without risk in any case where the tumour blood supply is derived exclusively or predominantly from branches of external carotid artery.

Adult↗

A DNA fragment from the origin of single-strand to double-strand DNA replication of bacteriophage fd.

A specific complex is formed between fd DNA, Escherichia coli DNA unwinding protein, and RNA polymerase (EC 2.7.7.6; nucleosidetriphosphate:RNA nucleotidyltransferase) during the first steps in the conversion of the single-stranded viral DNA to the double-stranded replicative form. In this complex a unique DNA fragment of about 120 nucleotides is protected against nuclease digestion. Both the requirements for its isolation and its position on the map of the phage genome indicate that the fragment contains the origin of single-strand to double-strand DNA replication. The isolated DNA fragment possesses double-strand-like characteristics, which protect it from being covered by the DNA unwinding protein and thus indirectly positions the RNA polymerase to the origin of replication.

Chromosome Mapping↗

[The aetiology and management of the frontal sinus mucopyocoele (author's transl)].

The pathogenesis of frontal sinus mucopyocoele is assessed in 33 patients seen between 1958 to 1972. In 45% there has been previous frontal sinus surgery. The importance of the frontonasal duct and healthy mucosa is stressed. The unexpectedly high number of mucopyocoeles following surgery permitted critical assessment of the procedures employed. The most successful and least complicated of which involved preservation or reconstruction of a drainage canal, removal of the obstructing middle turbinate and use of an indwelling drainage tube for at least 6 months. Regular follow-up and adequate early treatment of intercurrent infection are essential.

Frontal Sinus↗

Stabilization of promoter complexes with a single ribonucleoside triphosphate.

Under specific binding conditions RNA polymerase forms complexes at several sites of the replicative form DNA of bacteriophage fd. One of these complexes becomes stable to both high salt and low temperature after incubation with GTP. None of the complexes is stabilized by ATP. The stabilization by GTP results from the synthesis of an oligo(G) chain, which is bound in the complex. Size and pyrimidine fingerprints of the DNA segment protected by the enzyme against digestion with DNase are not changed upon initiation of oligo(G) synthesis. This result indicates that binding site and initiation site are identical parts of a promoter region.

Binding Sites↗

Initiation of transcription within an RNA-polymerase binding site.

1. The 5'-terminal sequence of the RNA transcribed from bacteriophage fd replicative form DNA under the control of promotor region I has been determined to be ppp(Gp)nUpApApApGpApCpCpUpGpApUpUp. . . 2. This sequence is complementary to the 5'-terminal sequence of the minus strand of the corresponding RNA polymerase binding site I, the starting point for RNA synthesis lying approximately in the middle of the binding site. 3. This initial sequence is also transcribed faithfully from isolated complexes of RNA polymerase and binding site I, obtained by DNase digestion of complexes between RNA polymerase and fd replicative form DNA. These highly stable complexes can not be reconstituted from binding site and enzyme. 4. It is concluded that RNA polymerase binding site and initiation site are identical parts of a promoter region, and that no "drift" between these sites is required as a step in RNA chain initiation. An additional non-transcribed outside region is implicated as essential for full promoter function.

Base Sequence↗

Nucleotide sequence of an RNA polymerase binding site from the DNA of bacteriophage fd.

The primary structure of a strong RNA polymerase binding site in the replicative form DNA of phage fd has been determined by direct DNA sequencing. It is: (see article). The molecule contains regions with 2-fold symmetry and sequence homologies to promoter regions from other DNAs. The startpoint of transcription is located in the center of the binding site.

Base Sequence↗

Isolation and characterization of thermosensitive Escherichia coli mutants defective in deoxyribonucleic acid replication.

Thermosensitive deoxyribonucleic acid replication-defective mutants have been isolated by using an autoradiographic selection method. The mutants have been analyzed genetically and biochemically. Some of the mutants show thermosensitivity of in vitro deoxyribonucleic acid replication. These can be classified into three groups according to their behavior in in vitro complementation assays. This classification is congruent with that obtained by genetic mapping by using cotransduction frequencies with selected markers in P1 transduction analysis.

Autoradiography↗