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Biomedical subjects

J Nüesch

Publications and source records attributed to J Nüesch.

At least 19 recordsLinked to original sources
PubMed1988

Detection and characterization of subgenomic RNAs in hepatitis A virus particles.

Defective viral particles containing deleted genomes were detected in harvests of cell cultures infected with various HAV isolates. The most prominent deletions were identified within the region of the genome coding for structural proteins. In this location three different deletions spanning nts 930-4380 (A), 1140-3820 (B), and 1370-3240 (C) were characterized. In addition to these internal deletions, various truncated RNAs were detected lacking either partially or completely the 3' terminal region which is supposed to code for viral replicase. RNA molecules with internal deletions as well as those with 3' terminal truncations could also be extracted directly from infected cells. During multiple consecutive passages of a given HAV strain, deletions A, B, and C accumulated and a quantitative increase of deleted RNAs occurred. Type and predominance of deletions varied with virus strains (CLF, GBM, MBB11/5, HM175, CR326, H141) and with the type of cells used for propagation (MRC-5, BGM, HELF, PLC/PRF/5). However, within the limits of the reliability of S1 analysis the endpoints of deletions A, B, and C were conserved. The mechanisms leading to formation of deletions remain unclear. Yet, some sequences flanking internal deletions showed homology with common splice signals and 3' terminal truncations proved to be confined to a distinct region within the genome.

Capsid↗
PubMed1984

Structural and functional organization of the gpt gene region of Escherichia coli.

The nucleotide sequence was determined for the Escherichia coli region containing the gpt gene, which encodes the enzyme xanthine-guanine phosphoribosyl transferase (XGPRT; EC 2.4.2.22). Restriction enzyme and sequence analyses have allowed us to locate precisely the gpt gene (16.9-kDal XGPRT) with respect to other genes in this region, notably phoE. Genes gpt and phoE are pointing towards each other and are separated by about 1840 bp. Available sequence data and protein analyses [Overbeeke et al., J. Mol. Biol. 163 (1983) 513-522, and this paper] indicate the presence, between gpt and phoE, of two additional genes. These genes are oriented the same way as gpt and code for proteins of 49 and 15.7 kDal, respectively. By in vitro transcription with E. coli RNA polymerase and nuclease S1 analysis, we have identified a promoter upstream of gpt. The short intercistronic region between gpt and the 49-kDal protein gene contains a rho-independent termination signal that closely precedes and partially overlaps another promoter. It appears from these data that gpt transcription is essentially monocistronic, giving rise to RNA of approx. 555 nucleotides, whereas the 49-kDal and 15.7-kDal protein genes are transcribed from their own promoter.

Amino Acid Sequence↗
PubMed1984

Partial purification and catalytic properties of a bifunctional enzyme in the biosynthetic pathway of beta-lactams in Cephalosporium acremonium.

The catalytic properties of the partially purified deacetoxycephalosporin C (DAOC)-synthetase and DAOC-hydroxylase from an industrial strain of Cephalosporium acremonium were studied. After mechanical breakage of the cells, purification was achieved by fractional (NH4)2SO4 precipitation, gel chromatography on Sephadex G-75, ion exchange chromatography on DEAE-Trisacryl M and two isoelectric focusing steps. The two enzyme activities could not be separated. Indirect evidence was obtained from SDS-polyacrylamide gel electrophoresis of the purest fractions obtained by isoelectric focusing that the two reactions are catalyzed by a single enzyme with a molecular weight of 33,000 +/- 2,000 and a pI of 4.6 +/- 0.1. Both reactions require alpha-ketoglutarate, FeSO4, ascorbate and O2, whereas additional ATP shows only a slight stimulation.

Acremonium↗
PubMed1978

[What is the contribution of colposcopic polypectomy?].

Coloscopic polypectomy saves patients with polyps of the colon from diagnostic laparotomy and also serves for the diagnosis and in the majority of patients even therapy of early colonic cancer. Complications are rare and most can be controlled by endoscopy. Recurrences of the polyp are possible, and therefore, follow-up is necessary.

Aged↗
PubMed1978

A genetic approach to the biosynthesis of the rifamycin-chromophore in Nocardia mediterranei. I. Isolation and characterization of a pentose-excreting auxotrophic mutant of Nocardia mediterranei with drastically reduced rifamycin production.

The mutant under study, designated A8, is derived from a Nocardia mediterranei strain, N813, which is a high rifamycin B producer. A8 is auxotrophic for aromatic amino acids and produces much less rifamycin B than the parent. A mixture of pentoses with D (--) ribulose as the main product is accumulated in the fermentation broth of this mutant. It was shown to be affected in its transketolase activity as no formation of D-sedoheptulose -7P from pentose-phosphates could be detected in vitro using crude extracts. The only pathway so far known which is derived from D-sedoheptulose-7P is the shikimate pathway leading to aromatic amino acids and vitamins. Biochemical and genetic investigations with mutant A8, which is defective in both the biosynthesis of rifamycins and the biosynthesis of shikimate pathway products, show that the seven-carbon amino unit of the rifamycin-chromophore must be derived from an intermediate of the shikimate pathway.

Culture Media↗
PubMed1978

A genetic approach to the biosynthesis of the rifamycin-chromophore in Nocardia mediterranei. II. Isolation and characterization of a shikimate excreting auxotrophic mutant of Nocardia mediterranei with normal rifamycin-production.

The mutant under study, designated A10, is derived from a Nocardia mediterranei strain, N813, which is a high rifamycin B producer. A10 is auxotrophic for aromatic amino acids but unlike A8 (see preceding paper) produces the same amount of rifamycin B as the parent. Shikimic acid and 3-dehydroshikimic acid are accumulated in the fermentation broth of this mutant. It was shown to be blocked in one of the enzymes leading from shikimate to chorismate. No formation of shikimate-3-phosphate from shikimate and ATP could be detected in vitro using crude extracts of this mutant and of the parent. As mutant A10 is only defective in the biosynthesis of aromatic amino acids and not in the biosynthesis of rifamycins it would appear that the seven-carbon amino unit of the rifamycin-chromophore must be derived from an intermediate of the shikimate pathway not behind shikimate. By referring to the results of the preceding paper it can be seen that the origin of this moiety can definitely be localized between 3-deoxy-D-arabinoheptulosonic acid-7-phosphate and shikimate.

Culture Media↗
PubMed1976

Enzymatic hydrolysis of cephalosporin C by an extracellular acetylhydrolase of Cephalosporium acremonium.

Extracellular hydrolases from Cephalosporium acremonium were analyzed according to their ability to deacetylate the beta-lactam antibiotic cephalosporin C. One out of at least six hydrolases exhibits appreciable cephalosporin C acetylhydrolase (CAH) activity. This enzyme was separated from other hydrolases and purified 220-fold. The purified CAH has a relatively low affinity for cephalosporin C (K(m), 20 mM) and is strongly inhibited by diisopropylfluorophosphate and less markedly affected by fluoride. Addition of glucose, maltose, and sucrose to the culture broth suppresses CAH production, whereas glycerol and succinate have no effect. Verrucarin A prevented the enzyme from appearing in the medium, which indicates the necessity of protein synthesis for CAH formation. When 1-thio-d-glucose was added to the culture medium, the results suggested that this glucose analogue is able to inhibit CAH synthesis. Our data provide evidence for a regulation of CAH synthesis similar to the catabolite repression system in bacteria.

Acremonium↗