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T Urasawa

Publications and source records attributed to T Urasawa.

At least 19 recordsLinked to original sources
PubMed2001

Sequence analysis of genes encoding structural and nonstructural proteins of a human group B rotavirus detected in Calcutta, India.

Nucleotide sequences of RNA segments encoding structural proteins(VP4, VP6, and VP7) and nonstructural proteins(NSP1 and NSP3) of a human group B rotavirus CAL-1, which was detected in Calcutta, India, were determined and their relatedness with cognate genes of other group B rotaviruses was analyzed. The CAL-1 genes showed generally high sequence identities (more than 90%) to those of human group B rotavirus, adult diarrheal rotavirus (ADRV) in China, while identities with bovine, murine, and ovine viruses were considerably lower (58-73%). Among RNA segments analyzed, sequence identity of the VP6 gene was relatively high compared with other gene segments. In the CAL-1 VP7 sequence, many characteristics were shared by ADRV, but not by other animal group B rotaviruses. In contrast, VP4 and NSP3 of CAL-1 were single amino acid and 23 amino acids longer than those of ADRV strain, respectively, due to differences of a few nucleotides. These findings suggested that human group B rotaviruses CAL-1 and ADRV might have originated from a common ancestral virus distinct from animal group B rotaviruses reported so far, while some notable sequence differences indicated the distinct nature of these viruses.

3' Untranslated Regions↗
PubMed2000

New P serotype of group A human rotavirus closely related to that of a porcine rotavirus.

The VP7 and VP4 genes of two human group A rotavirus strains Mc323 and Mc345 with unique serologic and genomic properties, and isolated in Chiang Mai, Thailand, in 1989 [Urasawa et al. (1992) Journal of Infectious Diseases 166:227-234] were further characterized. The nucleotide and deduced amino acid sequences of the VP7 genes allowed the classification of both strains as serotype G9. The VP4 genes of both strains are 2,359 nucleotides in length and encode a protein of 775 amino acids like in most human rotaviruses. A comparison of the VP4 amino acid sequence of strain Mc323 with those of strain Mc345 and 24 human and animal rotaviruses representing 20 distinct VP4 genotypes reported to date showed that VP4 of Mc323 and Mc345 belong to genotype 19 previously reported for porcine rotavirus [Burke et al. (1994) Journal of General Virology 75:2205-2212]. To investigate the serological type (P serotype) of these VP4s, six reassortant viruses each containing a distinct VP4 gene characteristic of human rotaviruses and the VP7 gene of porcine rotavirus strain Gottfried (G4) were prepared, and antisera to these reassortants produced in rabbits. In neutralization tests, the P serotype of Mc323 was clearly differentiated from the five major P serotypes reported previously for human rotaviruses, suggesting that Mc323 and Mc345 represent a new human rotavirus P serotype tentatively called P11.

Amino Acid Sequence↗
PubMed1999

Analysis of human rotavirus G serotype in Bangladesh by enzyme-linked immunosorbent assay and polymerase chain reaction.

Distribution of human rotavirus G serotype was investigated by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) with faecal specimens obtained from children with diarrhoea in Bangladesh. By ELISA, subgroup and G serotype were determined for 59.5% and 28.6% of group A rotavirus-positive specimens respectively. However, of the 120 specimens, the G serotype of which was not determined by ELISA, serotype of the 112 specimens was typed by PCR. In total, G serotype was assigned for 95.2% of all the specimens, showing the highest rate of G4 (41.7%), followed by G1 (23.2%) and G2 (14.9%). Twenty-four specimens showed mixed types, such as G2 with G1, G8 or G9, or G1 with G4. These results indicate that PCR combined with ELISA is highly effective for G serotyping of rotavirus.

Bangladesh↗
PubMed1998

Serological and genomic characterization of human rotaviruses detected in China.

A total of 1,385 stool specimens were collected from children with diarrhea at two hospitals in Wuhan, Hubei Province, China, in 1994 and 1995, and screened for rotavirus by polyacrylamide gel electrophoresis of viral RNA. Group A rotavirus was detected with high frequency; 56.5% (87/154) and 40.8% (502/1,231) of the specimens collected in 1994 and 1995, respectively, were positive for rotavirus. Assignment of G serotype and P type (VP4 genotype) of group A rotavirus by ELISA with monoclonal antibodies and/or PCR, respectively, showed that strains of G2-P[4] and G1-P[8] specificity were predominant in 1994 and in 1995, respectively. In contrast, a single strain was found to have a P[9] type specificity, and no G4 strain was detected. Unusual combinations of RNA pattern-subgroup-G serotype-P type, such as long pattern-subgroup I-G1-P[8], short pattern-subgroup II-G3-P[4] and short pattern-subgroup I-G1-P[4], were detected in four specimens. Nucleotide sequences of the VP8* and/or NSP5 genes from two Chinese P[8] strains 470 and 582 and one Chinese P[9] strain 512 as well as five Japanese P[9] strains (K8, AU1, M318, 0264, and 0265) were determined and compared with the published sequences of the corresponding gene. In the phylogenetic tree of VP8* sequences of P[9] strains, which formed two clusters each having strain K8 or AU-1 as the representative strain, the Chinese P[9] strain was found in the cluster represented by AU-1, although it was most distantly related to other strains. While NSP5 sequences of human strains with P[9] specificity were related to simian and bovine strains, that of Chinese P[8] strains was most closely related to those of porcine strains. A single group C rotavirus (No. 208) was detected. Nucleotide sequences of its VP4, VP6, VP7, and NSP4 genes were very similar to those of group C human rotaviruses detected worldwide.

Amino Acid Sequence↗
PubMed1996

Sequence analysis of normal and rearranged NSP5 genes from human rotavirus strains isolated in nature: implications for the occurrence of the rearrangement at the step of plus strand synthesis.

We determined the nucleotide sequences of normal and rearranged NSP5 genes from the human rotavirus strains (Mc323 and Mc345, respectively) which had previously been isolated from Thai infants with diarrhea in the same epidemic season. While the two strains shared G serotype 9 specificity and subgroup I specificity and they showed a high level of overall genomic relatedness to each other, they exhibited different RNA profiles; a long pattern for Mc323 and a super-short pattern for Mc345. Their NSP5 sequences were more closely related to those of porcine rotaviruses than to those of human rotaviruses. Mc345 NSP5 gene was shown to have concatemerization. In a head-to-tail orientation resulting in its length being 1182 bp, as compared with the Mc323 NSP5 sequence which was 664 bp in length. Sequence analysis suggested that the rearrangement found in the strain Mc345 occurred at the step of plus strand synthesis.

Base Sequence↗
PubMed1996

Structure and function of rotavirus NSP1.

Studies on the structure and function of the nonstructural proteins (NSP1-NSP5) of rotaviruses are important for dissection of the morphogenesis and replication processes of rotavirus. Above all, NSP1, the product of gene 5, has several interesting features, such as extreme sequence diversity, a highly conserved cysteine-rich region, RNA-binding activity, accumulation on the cytoskeleton, and non-random segregation in reassortment. Recently, comparable NSP1 sequence analysis has been performed on a number of rotavirus strains from various species. Furthermore, characterization of mutants with rearranged NSP1 genes has helped to elucidate the structure-function interaction of NSP1. We isolated and characterized two interesting mutants which have a large deletion including the cysteine-rich region or a nonsense codon at the early portion in the open reading frame (ORF) of the NSP1 gene. In this report, we summarize the structure and function of NSP1.

Amino Acid Sequence↗
PubMed1996

G (VP7) serotype-dependent preferential VP7 gene selection detected in the genetic background of simian rotavirus SA11.

We previously found the preferential selection of VP7 gene from a parent rotavirus strain SA11 with G serotype 3 (G3) in the sequential passages after mixed infection of simian rotavirus SA11 and SA11-human rotavirus single-VP7 gene-substitution reassortants with G1, G2, or G4 specificity. However, it has not been known whether or not VP7 genes derived from other strains with G3 specificity (G3-VP7 gene) are preferentially selected in the genetic background of SA11. To address this question, mixed infections followed by multiple passages were performed with a reassortant SA11-L2/KU-R1 (SKR1) (which possesses VP7 gene derived from G1 human rotavirus KU and other 10 genes of SA11 origin) and one of the five G3-rotaviruses, RRV, K9, YO, AK35, and S3. After the 10th passage, selection rates of SA11-L2/KU-R1 gene 9 (G1-VP7 gene) and gene 5 (NSP1 gene) reduced considerably (0 to 20.4%) in the clones obtained from all the coinfection experiments, while all or some of other segments were preferentially selected from SKR1 depending on the pairs of coinfection. When viral growth kinetics was examined, SKR1 exhibited better growth and reached a higher titer than any G3 viruses. Although the generated reassortants with VP7 gene and NSP1 gene derived from G3 viruses showed almost similar growth kinetics to that of SKR1 during the first 20 h of replication, the titers of these reassortants were higher than that of SKR1 after 36 h postinfection. The results obtained in this study suggested that G3-VP7 gene is functionally more adapted to the genetic background of SA11.

Animals↗
PubMed1996

Selection of rotavirus VP7 gene in the genetic background of simian rotavirus SA11: implications for rotavirus reassortant vaccine development.

We previously reported that the VP7 gene from simian rotavirus SA11 with G-serotype 3(G3-VP7 gene) was preferentially selected in the genetic background of SA11 compared with the G1- or G2-VP7 gene. In the present study, selection of the G4-VP7 gene in competition with G1-, G2- or G3-VP7 gene in the SA11 background was analyzed through mixed infection experiments using SA11 and SA11-human rotavirus single-VP7 gene-substitution reassortants with G-serotypes 1, 2, and 4 (G1-, G2- and G4-reassortant). In virus clones from coinfection of SA11 and G4-reassortant, the frequency of G4 virus decreased to 7% at the 3rd passage and the G4 virus disappeared at the 10th passage, whereas the majority of the clones possessed G3 specificity. However, the predominance of either of the viruses coinfected was not observed in the mixed infection with G4-reassortant and G1- or G2-reassortant. Although growth kinetics of SA11 and G4-reassortant was similar, G4-reassortant showed significantly smaller plaque size than SA11, G1- and G2-reassortant did. These results indicated that the G3-VP7 gene from SA11 might be preferentially selected in the SA11 genetic background compared with the G4-VP7 gene, and suggested that the introduction of a single G4-VP7 gene may affect growth characteristics of recipient virus SA11. These results together with our previous findings suggested the significance of genetic compatibility between recipient viral genes and foreign VP7 gene in the development of multivalent reassortant rotavirus vaccines.

Animals↗
PubMed1996

The distribution of G and P types within isolates of bovine rotavirus in Japan.

Various combinations of G type and P type were observed in 76 bovine rotavirus (BRV) strains isolated from 235 diarrheal calves in three prefectures of Japan in 1992-1994. The most prevalent combination was G6:P5 (46/76, 60.5%), followed by G10:P11 (13/76, 17.1%), G6:P11 (7/76, 9.2%) and G10:P5 (5/76, 6.6%). No G6:P1 strain of BRV was recognized from the isolates in the present study, though this type of BRV is well known as a suitable vaccine strain against BRV infection.

Animals↗
PubMed1996

Serological and genomic characterization of porcine rotaviruses in Thailand: detection of a G10 porcine rotavirus.

A total of 557 fecal specimens collected from piglets with diarrhea in Thailand were examined for rotavirus RNA by polyacrylamide gel electrophoresis. Twenty-three, one, and two samples were positive for group A, group B, and group C rotaviruses, respectively. Two samples exhibited two segments found in picobirnavirus RNA. RNA electropherotyping of 23 group A rotaviruses showed that they were classified into five patterns. By serotyping by enzyme-linked immunosorbent assay and PCR, viruses in 3 and 14 specimens were found to be serotype G3 and serotype G10, respectively. For one specimen, containing a serotype G10 virus (strain P343), virus was isolated in MA-104 cells, and the nucleotide sequences of the VP7 and VP4 genes were determined. Comparative sequence analysis and cross-neutralization tests showed that strain P343 has B223-like G10 and UK-like P7 serotype (or VP4 genotype 5) specificities. Rotaviruses having such antigenic specificities have not been detected in piglets. Thus, the interspecies transmission of rotaviruses between cows and pigs was suggested.

Amino Acid Sequence↗
PubMed1995

Preferential selection of VP7 gene from a parent rotavirus strain (SA11) in sequential passages after mixed infection with SA11 and SA11-human rotavirus single-VP7 gene-substitution reassortants.

We studied the competitive growth among SA11-L2(G3) and its single-human VP7 gene-substitution reassortants SA11-L2/KU-R1(G1) and SA11-L2/DS1-R1(G2), which have the genetic background of SA11-L2, during sequential passages after mixed infection. When the same infectious units (m.o.i. of 5 p.f.u./cell) of SA11-L2 and a reassortant SA11-L2/KU-R1 were inoculated onto and passaged in MA104 cells, 88% of the virus clones isolated from the culture fluid at the 3rd passage belonged to G3, and all the clones from the 10th passage had G3 specificity. Even when SA11-L2/KU-R1 with titer 10 times higher than that of SA11-L2 was used in the coinfection, the predominance of clones with G3-VP7 was observed. Although G2 clones slightly surpassed G1 clones in number in the mixed culture of SA11-L2/KU-R1 and SA11-L2/DS1-R1, G3 clones predominated in the virus progeny from a mixed culture infected with the same titers of SA11-L2, SA11-L2/KU-R1, and SA11-L2/DS1-R1. However, no significant difference in viral growth was detected among SA11-L2 and the two reassortants.

Antigens, Viral↗
PubMed1995

Persistence of a single electropherotype and serotype (G6P5) of bovine rotavirus in calves on a closed dairy farm from 1990 to 1993.

A virologic survey was conducted on calves with diarrhea associated with bovine rotavirus (BRV) on a closed dairy farm. The BRV was detected from 32 of 219 (14.6%) fecal specimens repeatedly collected from 56 calves born during the years 1992-1993, regardless of whether they had diarrhea. Most of the 32 strains were isolated from fecal specimens obtained from 2- to 6-week-old calves. After electrophoresis of double-stranded viral RNA from the 32 strains, genomic RNA migration patterns were similar to those of the predominant BRV strains isolated at the same farm during the years 1990-1991. All representative strains were identified as G serotype 6 (G6) and P type 5 (P5) by results of the virus-neutralization test and polymerase chain reaction procedure. Thus, BRV had no change in genomic RNA electropherotypes and serologic antigenicities in a closed dairy herd over a period of several years.

Animals↗
PubMed1994

Species specificity and interspecies relatedness in VP4 genotypes demonstrated by VP4 sequence analysis of equine, feline, and canine rotavirus strains.

We determined the nucleotide and deduced amino acid sequences of the VP4 genes of five equine, two feline, and two canine rotavirus strains. A high degree of homology (> 97.0%) was found among the VP4 amino acid sequences of the equine strains H2, FI-14, and FI23. Equine strain L338 has a distinct VP4 amino acid sequence from those of the other equine strains (78.1% or less homology), and the L338 VP4 exhibited more than 17.0% divergence at the amino acid level from those of rotavirus strains published so far. The VP4 amino acid sequence of equine strain H1, which showed low homology with those of other equine strains, shares > 95.4% homology to those of porcine strains OSU and YM. VP4 amino acid sequences of feline strain Cat97 and canine strains CU-1 and K9 showed a high degree of homology (96.8 to 97.2%) to one another, and were found to be quite similar (96.0-97.0% homology) to that of a human HCR3 strain recently characterized. Feline strain Cat2, whose VP4 sequence is distinct from that of strain Cat97, has a VP4 similar to those of human strains K8 and AU-1 (97.8 and 97.5% homologies at amino acid level, respectively). Thus, the VP4 sequences of rotaviruses showed species specificity and interspecies relatedness.

Amino Acid Sequence↗
PubMed1994

Differences in plaque size and VP4 sequence found in SA11 virus clones having simian authentic VP4.

We isolated five stable SA11 clones (TN-S1, TN-S2, TN-L1, TN-L2, and BN-S4) with different plaque sizes from two SA11 stocks. In polyacrylamide gel electrophoresis, the mobilities of the fourth, fifth, and seventh RNA segments of SA11 clones with large plaque size (TN-L1 and TN-L2) were faster than those of clones with small plaque size (TN-S1, TN-S2, and BN-S4). Nucleotide sequence determination of the fourth RNA segment identified a five-amino-acid difference in VP4s between the clones with large and small plaque sizes. The VP4 sequences of two clones with small plaque size (TN-S1 and BN-S4) were the same as the sequence of SA11-SEM reported by K. Nishikawa et al. (1988, J. Virol. 62, 4022-4026), while the VP4 sequences of the clones with large plaque size (TN-L1 and TN-L2) were similar, but not identical, to that reported by D. B. Mitchell and G. W. Both (1989, Nucleic Acids Res., 17, 2122). A single-gene reassortant, K8-L2.4, in which RNA segment 4 was derived from clone TN-L2 of SA11 virus and the other RNA segments were from strain K8, produced large plaques like TN-L2, suggesting that the five-amino-acid difference in VP4 between the clones with large and small plaque sizes might be associated with the difference in plaque size.

Animals↗
PubMed1994

Effect of the selection pressure with anti-VP7 and anti-VP4 neutralizing monoclonal antibodies on reassortant formation between two human rotaviruses.

In order to study the effect of selection pressure of anti-VP4 and anti-VP7 neutralizing monoclonal antibodies(N-MAbs) on reassortant formation, 424 reassortant clones were produced from mixed cultures of human rotavirus strains Wa and HN126 and their genotypes were analysed. Reassortant selection was done with four types of N-MAb: anti-VP4 MAb to Wa and anti-VP7 MAb to HN126(selection A); anti-VP4 MAb to HN126 and anti-VP7 MAb to Wa(selection B); anti-VP7 MAb to Wa(selection C); and anti-VP4 MAb to Wa(selection D). In each selection experiment, more than 100 clones were isolated, and the parental origin of RNA segments was identified by polyacrylamide gel electrophoresis. All clones isolated by selections A and B were found to be antigenic mosaic reassortants with the VP4 gene of HN126 and the VP7 gene of Wa and antigenic mosaic reassortants with the VP4 gene of Wa and the VP7 gene of HN126, respectively. Although in reassortants of both selections, RNA segments 2, 3, 5 and 6 were selected from strain Wa at considerably high rates, selection rates of RNA segments 1, 7, 8, and 11 were significantly different between selection A and B. In reassortants from selection C and D, selection rates of RNA segments 1, 3, 6, 7, 8, and 11 from Wa were significantly lower than those in selection A and B, whereas RNA segments 2 and 5 were almost exclusively selected from Wa as observed in selection A and B. These results indicated the presence of two types of nonrandom gene selection in reassortant formation, one strongly dependent on, and another irrespective of, the selection pressure with N-MAbs.

Antibodies, Monoclonal↗
PubMed1994

Survey on the distribution of the gene 4 alleles of human rotaviruses by polymerase chain reaction.

The presence of six gene 4 alleles (or VP4 genotypes) in human rotaviruses has been recognized. Using 16 representative cultivable human rotavirus strains, we confirmed the specificity of VP4 genotyping by polymerase chain reaction (PCR) using the nested oligonucleotides specific to each of the four representative gene 4 alleles. Using the PCR, we surveyed the gene 4 alleles of 199 human rotaviruses in stools collected in Japan and Thailand. Strains with the gene 4 allele, corresponding to P1A serotype, were shown to be the most prevalent, but two strains with P2 gene 4 allele and one strain with P3 gene 4 allele were detected in Thailand and in Japan, respectively.

Alleles↗
PubMed1993

Nucleotide sequence of VP4 and VP7 genes of a unique human rotavirus strain Mc35 with subgroup I and serotype 10 specificity.

The VP7 and VP4 genome segments of human rotavirus strain Mc35 (Urasawa et al., J. Infect. Dis. 166, 227-234, 1992) were investigated. Analysis of its VP7 nucleotide and amino acid sequences confirmed that it belongs to G (or VP7) serotype 10; however, the nucleotide sequence homologies between Mc35 and the four G10 bovine strains examined were lower than those among the four G10 bovine strains. The VP4 nucleotide and amino acid sequences of Mc35 were distinct (66.1-68.8% nucleotide and 64.9-70.0% amino acid homologies) from those of previously reported human and animal rotavirus strains representing different P (or VP4) types except for the strain K8 VP4, which was designated as type 3 of human rotavirus VP4 by Gorziglia et al. (Proc. Natl. Acad. Sci. USA., 87, 7155-7159, 1990). The VP4s of the K8 and Mc35 strains showed a relatively high degree of homology (77.6% nucleotide and 86.0% amino acid sequence homologies) and were presumed to represent subtypes of type 3 human rotavirus VP4 (type 3A (K8 VP4) and type 3B (Mc35 VP4)).

Amino Acid Sequence↗