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H Lother

Publications and source records attributed to H Lother.

28 records · Page 2Linked to original sources

Effect of dam methylation on the activity of the E. coli replication origin, oriC.

Methylation of GATC sites by the dam methylase is required for efficient initiation of DNA replication at the replication origin, oriC, of Escherichia coli. This is demonstrated by the inability of minichromosomes to be maintained in dam mutant strains. The requirement for methylated GATC sites is less stringent in vitro than in vivo. The time required for complete methylation of the origin region apparently determines the minimal spacing of replication forks on the chromosome.

Chromosomes, Bacterial↗

AsnC: an autogenously regulated activator of asparagine synthetase A transcription in Escherichia coli.

The regulation of the asparagine synthetase A gene of Escherichia coli was studied in vitro with a coupled transcription-translation system. It was shown that the 17-kilodalton gene, which is transcribed divergently from the adjacent asnA gene, codes for an activator of asnA transcription. The synthesis of the 17-kilodalton protein, which we now call AsnC, is autogenously regulated. The stimulating effect of AsnC on asnA transcription is abolished by asparagine, while the autoregulation of asnC is not affected by asparagine. The N-terminal part of the asnC protein, inferred from the DNA sequence, is homologous to the DNA-binding domain of regulatory proteins like catabolite gene activator, cro, and cI. This homology and direct repeats found in the region of the two asn promoters suggest that the asnC protein regulates transcription by binding to DNA. The asn promoters were defined by mapping of the mRNA start sites of in vitro-generated transcripts.

Asparagine↗

DNA binding and antigenic specifications of DNA gyrase.

Complexes of DNA gyrase and minichromosomal DNA containing the origin of replication of Escherichia coli (oriC) can be formed without metabolic energy and visualised by electron microscopy. The A subunit, part of the A2B2-DNA gyrase complex is the binding protein. Various binding sites are scattered around the minichromosomal DNA including oriC. The minimal origin contains the only prominent and reproducible binding site. Binding to this site is suppressed by oxolinic acid and the ATP analogue beta-y-imido ATP. If gyrase isolated from the gram-positive bacterium Bacillus subtilis is used no binding to oriC is seen. This observation is consistent with antigenic differences between the A subunits of the two microorganisms. The binding to oriC might reflect a requirement for DNA gyrase during the initiation of DNA replication.

Antigen-Antibody Complex↗

Recognition sites for a membrane-derived DNA binding protein preparation in the E. coli replication origin.

The DNA binding protein B' preparation, isolated from the membrane of E. coli, recognizes two sites, one of which is located in the minimum oriC (35-270 bp) and the other between base pairs 417 and 488. Recognition is only possible when restriction fragments containing these sites are in single-stranded state. At the first site the strand reading 3'OH-5'P in the direction of the E. coli genetic map is recognized, at the second site the 5'P-3'OH strand.

Bacterial Proteins↗

Purification of the E. coli dnaA gene product.

The product of the dnaA gene of Escherichia coli was isolated in a highly enriched form. The purification product binds specifically to DNA containing the E. coli chromosomal origin of replication, oriC.

Bacterial Proteins↗

Studies on the relevance of microtubules and of microfilament-dependent processes for triggering lymphocyte activation.

The effects of Isoptin i) on isolated microtubules, ii) on the anti-immunoglobulin and the Concanavalin A induced changes in the plasma membrane topography of murine lymphocytes and iii) on murine lymphocyte activation by lipopolysaccharide and by Concanavalin A was investigated. Isoptin and lidocaine (a local anaestetic) inhibited repolymerisation of tubulin into microtubules and induced depolymerisation of microtubules. Isoptin and lidocaine inhibited competitively binding of colchicine (a classical microtubules disrupting agent) to tubulin. Isoptin induced changes in the plasma membrane topography resembling effects caused by local anaesthetics or by a combination of colchicine and cytochalasin B (an agent affecting microfilament function). Isoptin, lidocaine, colchicine and hydroxyurea when present in the culture medium during the whole incubation period inhibited DNA synthesis induced by lipopolysaccharide or Concanavalin A. RNA synthesis was completely inhibited by lidocaine but not by Isoptin or by colchicine. If Isoptin, colchicine or hydroxyurea were removed from the culture medium at 20 h of culture period, the cells immediately started to incorporate 3H-Thymidine. The inhibitory action of lidocaine was irreversible. These results show that Isoptin, a drug which depolymerizes microtubules in vitro and disturbs the mitogen induced changes in plasma membrane topography of lymphocytes (believed to be controlled by microtubules and microfilaments), does not abolish commitment of the cells for DNA synthesis.

Animals↗

Distribution of membrane-bound and free ribosomes in growing yeast.

1. The distribution of membrane-bound and free ribosomes was investigated in stationary as well as in growing yeast cells. the relative amount of free ribosomes varies with the growth phase of the cell culture. During the duplication phases of the cell, relative maxima of free ribosomes can be found. However, the absolute amount of free ribosomes is fairly constant during the growth of the cells. 2. Membrane-bound ribosomes show lower polypeptide synthesis activity in a cell-free, poly (U)-dependent system than free ribosomes. 3. There is no difference in the distribution pattern of free and membrane-bound ribosomes in growing yeast cells of different ploidy. 4. A turnover between free and membrane-bound ribosomes is suggested to be in agreement with the hypothesis of Branes and Pogo ((1975) Eur. J. Biochem. 54, 317-328).

Binding Sites↗