PubMed Health⌕ Search

Biomedical subjects

R Rott

Publications and source records attributed to R Rott.

At least 91 records · Page 5Linked to original sources
PubMed1988

Chemical identification of cysteine as palmitoylation site in a transmembrane protein (Semliki Forest virus E1).

The palmitoylation site of the membrane glycoprotein E1 of Semliki Forest virus (SFV) has been identified by chemical analysis of an acylpeptide. 3H-Palmitoylated E1 isolated from SFV grown in baby hamster kidney cells was digested with chymotrypsin and the resulting peptides subjected to high performance liquid chromatography on a wide-pore column. The 3H-acylated peptide fraction peaked at above 60% 2-propanol in the eluent, indicating its hydrophobic character. Polyacrylamide gel electrophoresis analysis revealed a molecular weight of about Mr = 6000 for the radiolabeled peptide. Manual sequencing of this material by the 4-N,N'-dimethylaminoazobenzene-4'-isothiocyanate/phenylisothiocyanate procedure on solid phase revealed the amino-terminal sequence Ala-Ala-Ser-His-Ser-Asn-Val-Val-Phe-Pro. The same peptide also labels with [35S]cysteine. Comparison with the deduced amino acid sequence of E1 revealed that the palmitoylated peptide contains at least 43 amino acid residues, and thus includes the membrane spanning region down to the only cysteine residue five positions up from the carboxyl terminus of E1. Since [3H]palmitic acid was cleaved from E1 with thiol reagents, and since the peptide labels with [14C]iodoacetamide only after the release of fatty acids by hydroxylamine treatment, cysteine in position 433 represents the palmitoylation site in SFV E1.

Amino Acid Sequence↗
PubMed1988

Inhibition of glycoprotein oligosaccharide processing in vitro and in influenza-virus-infected cells by alpha-D-mannopyranosylmethyl-p-nitrophenyltriazene.

The effects of alpha-D-mannopyranosylmethyl-p-nitrophenyltriazene (MMNT) on mannosidases involved in asparagine-linked oligosaccharide processing were investigated. MMNT was found to inhibit the activity of rat liver Golgi alpha-mannosidase I in a concentration-dependent manner (50% inhibition with 0.18 mM-MMNT), whereas rat liver endoplasmic-reticulum alpha-mannosidase appeared to be resistant (less than 5% inhibition at 1 mM-MMNT). Jack-bean alpha-mannosidase was also sensitive to inhibition by MMNT (50% inhibition with 0.32 mM-MMNT). Treatment of influenza-virus-infected chick-embryo cells with 1 mM-MMNT led to a decrease in the formation of complex-type asparagine-linked oligosaccharides and an accumulation of high-mannose-type oligosaccharides with the composition Man8(GlcNAc)2 and Man7(GlcNAc)2 on the viral glycoproteins. The biological activities of influenza-virus haemagglutinin and neuraminidase synthesized in the presence of 1 mM-MMNT remained unchanged, but the virus was less infectious than the control.

Animals↗
PubMed1988

Animal viruses are able to fuse with prokaryotic cells. Fusion between Sendai or influenza virions and Mycoplasma.

Sendai and influenza virions are able to fuse with mycoplasmata. Virus-Mycoplasma fusion was demonstrated by the use of fluorescently labeled intact virions and fluorescence dequenching, as well as by electron microscopy. A high degree of fusion was observed upon incubation of both virions with Mycoplasma gallisepticum or Mycoplasma capricolum. Significantly less virus-cell fusion was observed with Acholeplasma laidlawii, whose membrane contains relatively low amounts of cholesterol. The requirement of cholesterol for allowing virus-Mycoplasma fusion was also demonstrated by showing that a low degree of fusion was obtained with M. capricolum, whose cholesterol content was decreased by modifying its growth medium. Fluorescence dequenching was not observed by incubating unfusogenic virions with mycoplasmata. Sendai virions were rendered nonfusogenic by treatment with trypsin, phenylmethylsulfonyl fluoride, or dithiothreitol, whereas influenza virions were made nonfusogenic by treatment with glutaraldehyde, ammonium hydroxide, high temperatures, or incubation at low pH. Practically no fusion was observed using influenza virions bearing uncleaved hemagglutinin. Trypsinization of influenza virions bearing uncleaved hemagglutinin greatly stimulated their ability to fuse with Mycoplasma cells. Similarly to intact virus particles, also reconstituted virus envelopes, bearing the two viral glycoproteins, fused with M. capricolum. However, membrane vesicles, bearing only the viral binding (HN) or fusion (F) glycoproteins, failed to fuse with mycoplasmata. Fusion between animal enveloped virions and prokaryotic cells was thus demonstrated.

Acholeplasma laidlawii↗
PubMed1988

Axonal transport of Borna disease virus along olfactory pathways in spontaneously and experimentally infected rats.

In this study it has been shown that infection of mother rats by Borna disease virus (BDV) from infected newborns led to a fatal disease. This differed both in clinical symptoms and in histological alterations from the form of the disease which occurred after intracerebral (i.c.) infection. Both parameters were, however, similar to those seen after experimental intranasal (i.n.) infection of adult rats. Detailed immunohistological studies showed clearly that after experimental i.n. infection, the infecting virus migrates intraaxonally from the neuroreceptors in the olfactory epithelium into the brain. It is therefore suggested that i.n. transmission is an important route of natural BDV infection.

Animals↗
PubMed1988

The structure of serotype H10 hemagglutinin of influenza A virus: comparison of an apathogenic avian and a mammalian strain pathogenic for mink.

The primary structure of the hemagglutinin of the apathogenic avian influenza virus A/chick/Germany/N/49 (H10N7) and of the serologically related strain A/mink/Sweden/84 (H10N4) pathogenic for mink has been elucidated by nucleotide sequence analysis, and the carbohydrates attached to the polypeptide have been determined. The H10 hemagglutinin has 65, 52, 46, 45, and 44% amino acid sequence homology with serotypes H7, H3, H1, H2, and H5, respectively. H10 and H7 hemagglutinins are also most closely related in their glycosylation patterns. There is a high sequence homology between both H10 strains supporting the concept that the mink virus has obtained its hemagglutinin from an avian strain. The sequence homology includes the cleavage site which consists of a single arginine as is the case with most other hemagglutinins exhibiting low susceptibility to proteolytic activation. The similarity in hemagglutinin structure between both H10 strains is discussed in light of the distinct differences in the pathogenicity of both viruses.

Amino Acid Sequence↗
PubMed1988

Characterization of a pantropic variant of Sendai virus derived from a host range mutant.

A variant (F1-R) was isolated from a temperature-sensitive host range mutant (ts-f1) of Sendai virus. F1-R was no longer temperature-sensitive but it retained the host range phenotype. Unlike wild-type virus, F1-R and ts-f1 undergo multiple cycles of replication in several cell lines in the absence of trypsin. This was attributed to proteolytic activation of the fusion (F) glycoprotein of the host range mutants, in cell nonpermissive to wild-type virus. In mice infected intranasally the variant F1-R caused a generalized infection. This was shown by immunohistology and with infectious virus being recovered from several organs whereas infection with wild-type virus was restricted to the lung. These observations indicate that the pantropic property of F1-R is the result of proteolytic activation of the virus by ubiquitous proteases. Nucleotide sequence analyses revealed that ts-f1 and F1-R differed from the wild-type virus by mutations at the region of the cleavage site of F and at the glycosylation site of the F2 subunit. The findings indicated that these mutations are responsible for the increased cleavability of the F protein of ts-f1 and F1-R and therefore are important determinants for the pantropism of F1-R.

Animals↗
PubMed1988

Protection against virulent H5 avian influenza virus infection in chickens by an inactivated vaccine produced with recombinant vaccinia virus.

A cloned cDNA copy of the haemagglutinin (HA) gene of A/Chicken/Scotland/59 (H5N1) influenza virus has been expressed in vaccinia virus. This pox virus is poorly infectious or non-infectious for chickens. However, immunization of chickens with lysates of cell cultures infected with the recombinant vaccinia virus, that had been emulsified with adjuvant and which contained an estimated 0.5 microgram influenza HA, elicited a substantial neutralizing antibody response to influenza virus. Challenges of immunized and non-immunized adult chickens with virulent A/Chicken/Scotland/59 influenza virus showed that the immunized animals were highly protected while the non-immunized controls died. Immunized birds were also protected against infection with the recent virulent H5 avian influenza virus, A/Chicken/Pennsylvania/83 (H5N2).

Animals↗
PubMed1988

Hyaluronidase enhances cell fusion and synthesis of viral DNA during infection with caprine arthritis encephalitis virus.

Caprine arthritis encephalitis virus (CAEV) is a lentivirus which infects goats and causes chronic progressive arthritis after a prolonged incubation period. CAEV replicates productively in cultures of goat synovial membrane cells and causes cytopathic effects characterized by multinucleated giant cell formation. The enzyme hyaluronidase was found to accelerate this virus induced fusion of GSM cells. Hyaluronidase treatment also resulted in synthesis of increased levels of unintegrated viral DNA early after infection. However, there was no significant increase in viral RNA in the infected cells or in the amount of virus produced. These studies suggest that hyaluronidase facilitates the interaction of CAEV with the target cells. Further it suggests that only a few copies of viral DNA are required to achieve maximal levels of virus replication. Additional copies of viral DNA appear to be redundant not contributing to viral specific transcription or increased production of virus.

Animal Diseases↗
PubMed1988

Immune-mediated pathogenesis of Borna disease.

Borna disease is an endemic progressive encephalomyelitis of horses and sheep prevalent in central Europe. A wide variety of animal species, ranging from chickens to primates can be infected experimentally with the causative virus, which is only poorly characterized. Furthermore, BD virus-specific antibodies have been detected in sera and cerebrospinal fluids of psychiatric patients. Our studies on the pathogenesis of BD have shown that-at least in rats-the disease is not caused by the infecting virus itself, but by a virus-induced immunopathological reaction. Thus, after intracerebral infection immunoincompetent rats do not get the disease despite persistent virus replication in cells of the central nervous system. However, after adoptive transfer of immune cells from diseased rats, immunoincompetent rats exhibit full-blown BD. Recently, we have been successful in establishing a virus-specific T cell line of the helper/inducer phenotype (CD4+). This T cell was shown to play an important role in the pathogenesis of BD, suggesting that the disease is caused by a delayed type hypersensitivity reaction.

Animals↗
PubMed1987

Protective effect of antibodies to two viral envelope glycoproteins on lethal infection with Newcastle disease virus.

The protective effect of humoral immunity against lethal infection of chickens with Newcastle disease virus was studied. Chickens hatched from eggs laid by hens vaccinated with live attenuated Newcastle disease virus vaccine possessed antibody to various components of the virus, and were resistant to a challenge with a virulent strain of Newcastle disease virus which was 100 per cent fatal for the offspring of nonvaccinated hens. Passive administration of antiserum raised against whole virions provided susceptible chickens protection comparable to that seen in the birds with maternal antibody. When administered passively, both anti-HN serum with virus neutralizing activity, and anti-F serum with only marginal virus neutralizing activity significantly prolonged the survival of infected birds but failed to achieve the level of protection as afforded by the anti-whole NDV serum. The protection provided by the simultaneous presence of anti-HN and anti-F serum was significantly greater than that afforded by either alone and comparable to that of anti-whole NDV serum, indicating the complementary effect of anti-HN and anti-F antibodies not only in cell cultures as reported previously (19), but also in a natural host.

Animals↗
PubMed1987

Aggravation of pathogenicity of an avian influenza virus by adaptation to quails.

Influenza virus A/turkey/Ontario/7732/66 (H 5 N 9), which is highly pathogenic to chickens, is nonpathogenic to quails. After intratracheal or intramuscular inoculation of quails, virus replication was limited to the respiratory tract, genital organs, and pancreas. However, aggravation of the pathogenicity was achieved through adaptation only by several passages of lung homogenates in quails. The adapted virus caused a fatal generalized infection in quails as well as in chickens. The pathogenic change of the virus could not be explained by a change in the proteolytic cleavability of the hemagglutinin, because no difference was found in the cleavability between the original and the adapted viruses. The adapted virus formed larger plaques and grew a little faster than the original one in both chicken embryo and quail embryo cells. The faster multiplication of the adapted virus at the site of infection might be the reason for its change in pathogenicity. The original virus could circulate among quails by a direct contact transmission without causing disease. The shed virus, however, caused a fatal infection in chickens when they were kept in contact with the infected quails. The epidemiological significance of this observation is discussed.

Adaptation, Physiological↗
PubMed1987

Synergistic role of staphylococcal proteases in the induction of influenza virus pathogenicity.

Several strains of Staphylococcus aureus have been found to secrete proteases that activate infectivity of influenza virus by proteolytic cleavage of the hemagglutinin. The enzymes of the bacterial strains Wood 46 and M 86/86 have been characterized in some detail and were found to be serine proteases. In their substrate specificities and inhibitor sensitivities they proved to be similar to, but not identical with trypsin and plasmin. The hemagglutinin of an individual virus strain could be cleaved by the proteases of some but not all staphylococcal strains, and a given enzyme could cleave only some but not all hemagglutinins analyzed. When mice were coinfected intranasally with the appropriate strains of influenza virus and S. aureus, the hemagglutinin was readily activated allowing multiple cycles of virus replication in the lung. Under these conditions, the animals came down with a fatal disease exhibiting extended lesions in the lung tissue. In contrast, after infection with virus or bacteria alone, there were no significant pathological changes. When the staphylococcal strain did not contain a protease that was able to activate the hemagglutinin of the coinfecting virus strain, the animals did not exhibit disease. These observations demonstrate that coinfecting bacteria can play an essential role in the development of influenza pneumonia by providing a protease suitable for cleavage activation of the hemagglutinin.

Animals↗
PubMed1987

Significance of viral glycoproteins for infectivity and pathogenicity.

Disease resulting from virus infection is a complex event depending on the close interaction of viral and cellular factors. Through the application of biochemical and genetic methods, it is now possible to gain an insight into the molecular basis of these interactions. Thus, it has been shown that the glycoproteins of enveloped viruses play a central role in the initiation of infection. They are responsible not only for the adsorption of virions to cellular receptors, but are also for the entry of the genome into the cell by the fusion of viral envelopes with cellular membranes. Evidence is growing that the fusogenic glycoproteins are frequently activated by cellular proteases. The structure of the proteins at the cleavage site and the availability of a suitable protease are critical for tissue tropism, spread of the virus in the infected organism and, thus, for pathogenicity. This will be demonstrated here by the example of the haemagglutinin of influenza viruses.

Animals↗
PubMed1987

N-acetyl-9-O-acetylneuraminic acid, the receptor determinant for influenza C virus, is a differentiation marker on chicken erythrocytes.

Erythrocytes from chicken of different age were analysed for their agglutinability by influenza C virus, which has been shown recently to use N-acetyl-9-O-acetylneuraminic acid as a high-affinity receptor determinant for the attachment to cells. Only with birds not younger than six days complete agglutination of the erythrocytes was observed. The hemagglutination titer which was initially low reached its maximum value at the age of about 20 days. Sialic acid was isolated from erythrocytes, purified and analysed by colorimetry, thin-layer chromatography, high-performance liquid chromatography, and gas-liquid chromatography-mass spectrometry. The sialic acid content of erythrocytes from one-day old and adult chicken was 21 micrograms and 18 micrograms sialic acid/ml packed erythrocytes, respectively. While N-acetylneuraminic acid was the major type of sialic acid on erythrocytes from both one-day old and adult chicken, N-acetyl-9-O-acetylneuraminic acid was only detected on red blood cells from adult animals accounting for 30-40% of total sialic acid. These results indicate that N-acetyl-9-O-acetylneuraminic acid, in addition to serving as a receptor determinant for influenza C virus, represents a developmental marker on chicken erythrocytes.

Animals↗