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H Shibuta

Publications and source records attributed to H Shibuta.

At least 37 records · Page 2Linked to original sources
PubMed1985

Experimental parainfluenza virus infection in mice: growth and spread of a highly pathogenic variant of parainfluenza 3 virus in the mouse brain.

We had previously showed that following intracerebral inoculation of newborn mice, the 910 N and M strains of bovine parainfluenza 3 virus induce a non-lethal hydrocephalus and a lethal disease with marked thymic and splenic atrophy, respectively. Moreover, only the M virus was lethal for 2-week-old mice. In the present study, we demonstrate that the M virus multiplies and spreads in the mouse brain invading the thalamus, hypothalamus and brain stem beyond the ependyma whereas the 910 N virus causes only slight ependymitis. This growth and spread of M virus was blocked by passive immunization 3 days after infection. Mouse embryo brain cell cultures were infected with M and 910 N viruses, about 50 per cent became antigen-positive for M whereas only a small proportion of cells were positive for the 910 N virus. However, the latter did produce higher yeilds of infectious virus than M.

Animals↗
PubMed1984

Nucleotide sequence of a Sendai virus genome region covering the entire M gene and the 3' proximal 1013 nucleotides of the F gene.

We determined the sequence of the 2,138 nucleotides in the Sendai virus genome just following the 3' proximal 3,686 nucleotides which we had previously reported (Nucleic Acids Res. 11, 7317-7330, 1983). This covers the entire third gene of 1,173 nucleotides and the 3' proximal 1,013 nucleotides of the fourth gene. Like the NP and P+C genes, both the third and fourth genes start from consensus sequence R1 (3'-UCCCAC(or UA)UUUC) at the 3' end and the third gene terminates with consensus sequence R2 (3'-AUUCUUUUU) at the 5' end. The third gene was identified as M, and the deduced 348 amino acids indicated that the M protein is rich in basic residues and has hydrophobic domains near the C-terminal. The fourth gene, although sequencing is not complete yet, was identified as F, since a large open reading frame found in the gene contains the characteristic sequence of 20 amino acids located at the N-terminal of the F1 protein. Analyses of the amino acid sequence suggested that the structure of the F gene product is NH2-signal peptide-F2-F1-COOH.

Animals↗
PubMed1983

Sequence of 3,687 nucleotides from the 3' end of Sendai virus genome RNA and the predicted amino acid sequences of viral NP, P and C proteins.

The sequence of 3,687 nucleotides from the 3' end of the Sendai virus genome (Z strain) was determined by a molecular cloning technique followed by rapid sequence analysis. Two large open reading frames, one consisting of 1,572 nucleotides and the other of 1,704 nucleotides, were observed in the region, that is OP-1 and OP-2 from the 3' end of the genome. The amino acid sequences of the gene products were predicted from the observed sequence. Determination of amino acid compositions of viral proteins, P, HN, Fo, NP and M, led us to conclude that NP and P are the gene products of OP-1 and OP-2, respectively. An additional open reading frame consisting of 612 nucleotides (OP-3) was discovered in the 3' most proximal region of OP-2. The predicted product of OP-3 was considered to be viral non-structural protein C. The leader sequence of 51 nucleotides at the 3' terminal of the genome and consensus sequences at 3' and 5' ends of each gene for proteins NP and P were identified.

Amino Acid Sequence↗
PubMed1983

Neuraminidase activity and syncytial formation in variants of parainfluenza 3 virus.

By a sensitive fluorometric assay method, we could definitely demonstrate neuraminidase activity for two variants of parainfluenza 3 virus, M and SC, which were previously shown to have no detectable neuraminidase activity. The enzyme activities of these viruses were very similar to each other, showing a much lower catalytic rate, a much higher Km value, and a more acidic pH optimum than those of the virus variants of high neuraminidase activity, 910N, LT, and MR. M and SC viruses eluted from guinea pig erythrocytes very poorly, whereas 910N and LT viruses eluted readily. M virus required the aid of a bacterial neuraminidase for effective growth and plaque formation in MDBK cells, but the virus grew well and formed plaques in R66 and Vero cells without the enzyme. SC virus required no exogenous neuraminidase for growth in all of these cell types. Depending on cell type, SC virus induced slight to extensive syncytial formation which was greatly inhibited by exogenous neuraminidase. In contrast, M virus induced extensive syncytial formation in all these cells regardless of the presence or absence of exogenous neuraminidase, although development and disintegration of the syncytia were more or less retarded by the enzyme, especially in MDBK cells. These results indicate that M virus possesses highly potent inducibility of syncytial formation which is further fortified by being low in viral neuraminidase activity.

Adsorption↗
PubMed1982

Restricted viral RNA synthesis in establishment of persistent infection in Vero cells with a Sendai virus mutant.

It was previously shown that a temperature-sensitive mutant of Sendai virus, ts-23, readily establishes persistent infection in Vero cells at 37 C, a permissive temperature for growth of the mutant. In the present study, it was demonstrated that the virus yield from ts-23-infected Vero cells at 37 C began to decrease 48 to 72 hr postinfection, after an initial phase of high virus production. Before the decrease in virus production, the formation of viral nucleoprotein declined, although synthesis of all species of viral protein continued. It was suggested that the limited formation of viral nucleoprotein and the decrease in virus production were due to the restriction of viral RNA synthesis which began to occur early after infection in ts-23-infected cells at 37 C. The mutant has a temperature-sensitive defect in RNA polymerase activity and the temperature 37 C, used for establishment of persistent infection, would be a semi-permissive temperature for the RNA polymerase activity of the mutant. The ts-23 mutant interfered with the replication of the parental wild virus in Vero cells at 37 C.

Animals↗
PubMed1982

Experimental parainfluenzavirus infection in mice: fatal illness with atrophy of thymus and spleen in mice caused by a variant of parainfluenza 3 virus.

Intracerebral inoculation of the YN strain of parainfluenza 3 virus was found to induce an acute fatal illness characterized by marked thymic and splenic atrophy in newborn mice. Previously we showed that the YN strain contains three distinct plaque-type variants, LT, SC, and M. Of these, the M-type variant induced this fatal illness, whereas the other two variants induced hydrocephalus.

Animals↗
PubMed1981

Parainfluenza 3 virus: plaque-type variants lacking neuraminidase activity.

Virus clones lacking detectable neuraminidase activity (SC-YN and M-YN) as well as those possessing it (LT-910N and LT-YN) were isolated from bovine strains of parainfluenza 3 virus. LT-910N and LT-YN viruses produced large turbid plaques in MDBK cells, and SC-YN virus produced small clear plaques. Incorporation of a bacterial neuraminidase in agar overlay medium made SC-YN virus form large turbid plaques, whereas it made M-YN virus form large clear plaques. However, M-YN virus formed only pinhole plaques or no plaques in the absence of neuraminidase. The exogenous neuraminidase had little effect on the plaque formation of LT-910N and LT-YN viruses. M-YN virus induced extensive syncytial formation, and SC-YN virus produced less extensive syncytial formation. The exogenous neuraminidase enhanced replication of SC-YN and M-YN viruses and reduced syncytial formation by these viruses. The enzyme had little effect on replication and cytopathic effect of LT-910N and LT-YN viruses. The reason for these effects of the exogenous neuraminidase is discussed.

Animals↗
PubMed1980

Isolation and characterization of temperature-sensitive mutants of Sendai virus.

Sixteen temperature-sensitive mutants of Sendai virus were isolated from mutagenized stocks (10 mutants, designated numerically) and persistently infected cultures (6 mutants, designated alphabetically). Based on complementation tests, virion-associated activities, thermal inactivation, and viral RNA and hemadsorbing antigen synthesis as well as virion production in chick lung embryo cells at nonpermissive temperature, these mutants were divided into seven groups as follows. i) HANA group mutants (ts-5, -9, -10, -201), defective in hemagglutinin-neuraminidase protein, complementation group I. ii) F group mutants (ts-18, -108), defective in hemolytic and cell-fusing activity, complementation group II. iii) Ts-43, defective in RNA polymerase activity, complementation group III. iv) Ts-23, defective in RNA polymerase activity, interfered with the other mutants in complementation tests. v) Ts-25, defective in the incorporation of hemagglutinin-neuraminidase protein into the virion at the stage of virus assembly. vi) Ts-110, belongs to F group mutants on one hand, but is considered to carry another undetermined defect. vii) C group (carrier culture-borne group) mutants (ts-a, -b, -c, -d, -e, -f), defective lesion not yet determined and belong to neither complementation group I nor II. Assignment of mutants in groups iv), v), vi), and vii) to complementation groups could not be achieved.

Cell Line↗
PubMed1979

Characterization of bovine parainfluenza virus type 3.

Bovine parainfluenza virus type 3 (PIV-3) has a buoyant density of 1.197. The RNA of PIV-3, like that of Sendai virus, is a single continuous chain which lacks polyadenylic acid sequences and tends to self-anneal to a marked extent. It has a sedimentation coefficient of 42S and a molecular weight of 4.5 X 10(6), being slightly smaller than Sendai virus RNA (47S, 5.3 X 10(6)). PIV-3 has 5 main structural proteins, of which 2 are glycoproteins. The molecular weights of protein 1, protein 2, protein 3, glycoprotein 1, and glycoprotein 2 were estimated to be 79,000, 68,000, 35,000, 69,000, and 55,000, respectively. Protein 2 was suggested to be nucleocapsid protein.

Animals↗
PubMed1977

Polyadenylate in the virion RNA of mouse hepatitis virus.

Mouse hepatitis (MH) virus was grown in SR-CDF1-DBT, a mouse cell line, and purified by ammonium sulfate precipitation and by density gradient centrifugation. Extraction of RNA from purified virions with 1% SDS and sedimentation analysis of the RNA revealed a major 50S component and two minor components. Treatment of virions with phenol/chloroform also produced the 50S component, although its yield was lower. MH virion RNA can bind to a poly(U)-fiberglass filter, indicating that MH virion RNA contains poly(A). A poly(A)-like fragment was isolated by digestion with ribonuclease A [EC 3.1.4.22] and T1 [EC 3.1.4.8] and by DEAE-Sephadex column chromatography. Analysis of the fragment for base composition showed it to be an adenine-rich material. Its chain length was about 90 nucleotides, as determined by ion-exchange chromatography and gel electrophoresis.

Cell Line↗