Search PubMed⌕ Search

Biomedical subjects

H Moriuchi

Publications and source records attributed to H Moriuchi.

At least 73 records · Page 4Linked to original sources

Proteins and cis-acting elements associated with transactivation of the varicella-zoster virus (VZV) immediate-early gene 62 promoter by VZV open reading frame 10 protein.

Varicella-zoster virus (VZV) open reading frame 10 (ORF10) protein, the homolog of herpes simplex virus type 1 (HSV-1) VP16, is a virion-associated transactivator of the VZV immediate-early (IE) gene 62 (IE62) promoter. VP16 forms a complex with cellular factors (Oct1 and host cell factor [HCF]) and TAATGARAT elements (found in all HSV-1 IE promoter/enhancer sequences) to mediate stimulation of IE transcription. The VZV IE62 promoter also contains three TAATGARAT-like elements. Mutagenesis studies of the VZV IE62 promoter indicated that TAATGARAT-like elements contribute to transactivation of the VZV IE62 promoter by ORF10 protein. Other cis-acting elements such as GA-rich and cyclic AMP-responsive elements were also needed for full transactivation by ORF10 protein. In mobility shift assays, ORF10 protein formed a complex with either of two TAATGARAT-like elements that lack an overlapping octamer-binding motif (octa-/TAATGARAT) but not with a TAATGARAT element with an overlapping octamer-binding motif (octa+/TAATGARAT). In contrast, VP16 formed a high-affinity ternary complex with an octa+/TAATGARAT element and a low-affinity complex with octa-/TAATGARAT elements. Addition of antibodies to ORF10 protein, Oct1, or HCF disrupted the complexes, demonstrating that ORF10 protein interacts with Oct1 and HCF. These results suggest that transactivation of the VZV IE62 gene by ORF10 protein and HSV IE genes by VP16 require similar cellular proteins but distinct cis-acting elements.

Base Sequence↗

The RING finger domain of the varicella-zoster virus open reading frame 61 protein is required for its transregulatory functions.

Varicella-zoster virus (VZV) open reading frame 61 (ORF61) protein transactivates expression of VZV promoters. VZV ORF61 is functionally homologous to herpes simplex virus type 1 (HSV-1)-infected cell protein 0 (ICP0); however, amino acid sequence homology of these two proteins is limited to the RING finger domains, a recently identified sequence motif composed of cysteine and histidine residues, located in their amino-terminal regions. A carboxy-terminal truncation mutant of ICP0 (which retains the RING finger domain) was previously shown to act as a potent transrepressor and as a dominant-negative mutant in the presence of full-length ICP0. We have shown that the corresponding region of ORF61 has similar properties. Amino-terminal truncation mutants of VZV ORF61 and HSV-1 ICP0, which lack the RING finger domain, cannot transactivate VZV promoters; however, fusion of the amino portion of one protein to the carboxy portion of the other restored the transactivating activity of the full-length proteins. To further study the role of the ORF61 RING finger domain, cysteine or histidine residues of this domain were individually replaced with serine or aspargine, respectively. Each of these amino acid substitution mutants in the RING finger abolished the transactivating activity of the full-length ORF61 protein. Each of the substitutions also reduced the transrepressing and dominant-negative properties of a carboxy-terminal truncation mutant of ORF61. These results indicate that the RING finger domain is required for the transregulatory functions of ORF61.

Amino Acids↗

Varicella-zoster virus (VZV) virion-associated transactivator open reading frame 62 protein enhances the infectivity of VZV DNA.

Varicella-zoster virus (VZV) open reading frame (ORF) 62 protein (the homolog of herpes simplex virus type 1 (HSV-1) ICP4) and ORF10 protein (the homolog of HSV-1 VP16) are virion-associated transactivators. To investigate whether these proteins function during the initial stages of VZV infection, human melanoma cells were cotransfected with purified VZV DNA, devoid of any structural proteins, along with a plasmid expressing VZV ORF62 or ORF10 under the control of the human cytomegalovirus major immediate-early promoter. Expression of ORF62 enhanced the infectivity of VZV DNA up to 70-fold. In contrast, expression of ORF10 enhanced the infectivity of VZV DNA only threefold. These results show that high-level expression of ORF62 protein increases the probability that transfected VZV DNA will result in productive infection, suggesting that this virion-associated transactivator (ORF62) has a critical role in initiating infection.

Base Sequence↗

Varicella-zoster virus open reading frame 4 protein is functionally distinct from and does not complement its herpes simplex virus type 1 homolog, ICP27.

Varicella-zoster virus (VZV) open reading frame 4 (ORF4) encodes a putative immediate-early protein which is homologous to herpes simplex virus type 1 (HSV-1) ICP27 on the basis of gene location and similarity in amino acid sequence. In transient expression assays, however, ORF4 and ICP27 exhibit different properties. ICP27 alone has little activity on target plasmids, but it acts as a transactivator or a transrepressor in the presence of other HSV-1 transactivators. In contrast, ORF4 directly transactivates plasmids containing homologous or heterologous promoters and has no apparent transrepressing activity. To further illuminate the functional similarities and differences between ORF4 and ICP27, Vero cell lines which express ORF4 under the inducible metallothionein promoter were constructed. Cell lines expressing functionally active ORF4 protein upregulated the expression of transfected VZV target plasmids but were unable to efficiently complement HSV-1 ICP27 mutants. These results indicate that, despite structural similarities, VZV ORF4 and HSV-1 ICP27 behave differently in transient expression assays and may play different roles in virus replication.

Animals↗

[Oleic acid-induced PaO2 decrease model for primary screening of drugs for hypoxemia: effects of tranexamic acid and procaterol hydrochloride on the decrease in PaO2].

We constructed an oleic acid (OA)-induced PaO2 decrease model in guinea pigs. We then examined several basic conditions to establish the primary screening model to determine useful drugs for hypoxemia. Hartley strain guinea pigs were anesthetized with pentobarbital (25 mg/kg) and catheterized into the subclavian artery for blood sampling and for measuring blood pressure; they were also catheterized into the subclavian vein for drug administration. Then the animal was immobilized with pancuronium (0.1 mg/kg) and ventilated by a ventilator with room air. The following results were obtained: 1) there were no significant fluctuations of PaO2, PaCO2 and pH throughout the 11 sampling over a 2-hr period. Airway pressure and blood pressure also remained relatively constant. 2) Percentage of decrease in PaO2 by OA (15 microliters/kg) in the hyperventilated group was greater than that in the normally-ventilated group. 3) Increasing doses of 10, 15, 30 and 60 microliters/kg of OA resulted in dose-dependently lower values of PaO2. 4) Tranexamic acid (2 g/kg, i.p.) significantly prevented the decrease in PaO2 at 10 and 15 min after OA (15 microliters/kg) injection. 5) Procaterol hydrochloride (0.1 microgram/kg, i.v.) failed to inhibit the decrease in PaO2 by OA (15 microliters/kg). These results suggest that by using a suitable ventilation and OA dose, this model could be used as a primary screening model of drugs for hypoxemia and that tranexamic acid might be an effective drug for hypoxemia caused by a mechanism by which OA decreases PaO2.

Animals↗

Varicella-zoster virus open reading frame 10 protein, the herpes simplex virus VP16 homolog, transactivates herpesvirus immediate-early gene promoters.

The varicella-zoster virus (VZV) open reading frame 10 (ORF10) protein is the homolog of the herpes simplex virus type 1 (HSV-1) protein VP16. These are two virion tegument proteins that have extensive amino acid sequence identity in their amino-terminal and middle domains. ORF10, however, lacks the acidic carboxy terminus which is critical for transactivation by VP16. Earlier studies showed that VZV ORF10 does not form a tertiary complex with the TAATGARAT regulatory element (where R is a purine) with which HSV-1 VP16 interacts, suggesting that ORF10 may not have transactivating ability. Using transient-expression assays, we show that VZV ORF10 is able to transactivate VZV immediate-early (IE) gene (ORF62) and HSV-1 IE gene (ICP4 and ICP0) promoters. Furthermore, cell lines stably expressing ORF10 complement the HSV-1 mutant in1814, which lacks the transactivating function of VP16, and enhance the de novo synthesis of infectious virus following transfection of HSV-1 virion DNA. These results indicate that ORF10, like its HSV-1 homolog VP16, is a transactivating protein despite the absence of sequences similar to the VP16 carboxy-terminal domain. The transactivating function of the VZV ORF10 tegument protein may be critical for efficient initiation of viral infection.

Amino Acid Sequence↗

Varicella-zoster virus (VZV) open reading frame 61 protein transactivates VZV gene promoters and enhances the infectivity of VZV DNA.

The varicella-zoster virus (VZV) open reading frame 61 (ORF61) protein is the homolog of herpes simplex virus type 1 (HSV-1) ICP0. Both genes are located in similar parts of the genome, their predicted products share a cysteine-rich motif, and cell lines expressing VZV ORF61 are able to complement an HSV-1 ICP0 deletion mutant (H. Moriuchi, M. Moriuchi, H. A. Smith, S. E. Straus, and J. I. Cohen, J. Virol. 66:7303-7308, 1992). In transient expression assays, HSV-1 ICP0 is a transactivator alone and transactivates in synergy with another viral transactivator, ICP4. However, VZV ORF61 represses the activation by VZV-encoded proteins ORF62 (the homolog of ICP4) and ORF4. To further characterize the function of VZV ORF61 and its role(s) in regulation of viral gene expression, we performed transient expression assays using target promoters from VZV, HSV-1, and unrelated viruses. In the absence of other viral activators, VZV ORF61 transactivated most promoters tested. In addition, a cell line stably expressing VZV ORF61 complemented the HSV-1 mutant in 1814, which lacks the transactivating function of VP16. The cell line expressing VZV ORF61 enhanced the infectivity of HSV-1 virion DNA. Moreover, transient expression of VZV ORF61 also enhanced the infectivity of VZV DNA. These results indicate that VZV ORF61 can stimulate expression of HSV-1 and VZV genes at an early stage in the viral replicative cycle and that ORF61 has an important role in VZV gene regulation.

Animals↗

Varicella-zoster virus open reading frame 61 protein is functionally homologous to herpes simplex virus type 1 ICP0.

The varicella-zoster virus (VZV) open reading frame 61 (ORF61) protein is thought to be the homolog of herpes simplex virus type 1 (HSV-1) ICP0, based on gene location and limited amino acid homology. However, HSV-1 ICP0 trans activates HSV-1 genes, while VZV ORF61 protein trans represses the function of VZV trans activators on VZV promoters in transient expression assays. To investigate the functional relatedness of HSV-1 ICP0 and VZV ORF61 protein, we established Vero and MeWo cell lines which stably express VZV ORF61 under the control of a metallothionein promoter and performed complementation studies with an HSV-1 ICP0 deletion mutant (7134). Mutant 7134 is impaired for plaque formation and replication at a low multiplicity of infection in cell culture, but these defects were complemented by up to 200-fold in Vero cell lines expressing VZV ORF61. Likewise, the efficiency of plaque formation was improved by up to 100-fold in MeWo cell lines expressing VZV ORF61. A cell line expressing another VZV immediate-early gene product (ORF62) was unable to complement mutant 7134. HSV-1 mutants which are deleted for other HSV-1 immediate-early gene products (ICP4, ICP27) were unable to grow in VZV ORF61-expressing cell lines. These results indicate that, despite marked differences in their sequences and in effects on their cognate promoters in transient expression assays, VZV ORF61 protein is the functional homolog of HSV-1 ICP0.

Amino Acid Sequence↗

Neutralizing antibody response to cytomegalovirus in seropositive pregnant women.

The neutralizing (NT) antibody response to human cytomegalovirus (HCMV) in seropositive pregnant women was evaluated to establish the role of protective humoral immunity in HCMV infections (reactivation) during pregnancy. Complement fixation (CF) antibody titers increased significantly in 54 (0.62%) of 8,663 seropositive cases tested during gestation. However, titers of NT antibody, when tested in the presence of complement (C'), increased as gestation progressed and exceeded 1:500 in most of the cases in whom the CF antibody titer had shown earlier an insignificant increase. When NT antibodies were tested in the absence of C', insignificant increase in titers were found. The results suggest that the presence of high titer of NT antibody may play an important role in protective humoral immunity against reactivation of HCMV in pregnant women, and may also be effective in preventing intrauterine infection and modulating the severity of perinatal infection in offspring borne of HCMV carrier mothers.

Antibodies, Viral↗

Community-acquired influenza C virus infection in children.

To clarify the epidemiologic and clinical features of community-acquired influenza C infection in children, we took specimens throughout the year from a larger number of patients with acute respiratory illnesses in a pediatric clinic in Yamagata, Japan. During a 2-year survey, 20 strains of influenza C virus were isolated from 13,426 specimens. These isolates were recovered throughout the year. The ages of patients with influenza C virus isolates ranged from 2 months to 11 years and peaked at the age of 1 year. The clinical diagnosis of patients with influenza C virus infection included bronchitis in one child and pneumonia in four. Community-acquired influenza C infection in children can cause a variety of respiratory illnesses that cannot be clinically differentiated from those caused by other viruses.

Child↗

The herald waves of influenza virus infections detected in Sendai and Yamagata cities in 1985-1990.

The community surveillance of respiratory virus infections performed during 1985-1987 in Sendai and 1988-1990 in Yamagata has identified a total of five herald waves of influenza virus infections: A/H3N2 virus infections in 1985 and 1989, A/H1N1 virus infections in 1986 and 1988, and type B virus infections in 1989. To investigate the antigenic and genetic relationships between the herald wave and epidemic strains, influenza A/H1N1 viruses isolated during the 1986 and 1988 herald waves were compared with those isolated during the 1986-1987 and 1988-1989 epidemic seasons, respectively, utilizing hemagglutination inhibition tests with anti-hemagglutinin monoclonal antibodies and oligonucleotide mapping of total viral RNAs. The results showed that multiple variants differing in antigenic and genetic properties were cocirculating during the 1986 herald wave as well as during each of the two epidemics (only one strain was isolated in the 1988 herald wave). It was also observed that viruses which had the antigenic and/or genetic characteristics closely similar to those of the viruses circulating in the 1986 and 1988 herald waves, were isolated during the winters of 1986-1987 and 1988-1989, respectively.

Antibodies, Monoclonal↗

Hypolipidemic effects of guar gum and its enzyme hydrolysate in rats fed highly saturated fat diets.

The effects of guar gum and its enzyme hydrolysate as well as fructooligosaccharide on lipid metabolism were compared in rats fed high-fat diets employing lard or palm oil as dietary fat (25% in the diets). Guar gum and the enzyme hydrolysate greatly increased cecal volatile fatty acid contents to a similar extent. Fructooligosaccharide also increased the variable but to a lesser extent. Not only guar gum but also the hydrolysate and the oligosaccharide reduced serum cholesterol levels irrespective of dietary fat sources but to a lesser extent. The triglyceride-lowering effects of the hydrolysate and the oligosaccharide were comparable to that of guar gum. Although guar gum enzyme hydrolysate and fructooligosaccharide doubled the biliary bile acid excretion, these materials only slightly increased the activities of hepatic enzymes of cholesterol and bile acid synthesis. Guar gum and its hydrolysate suppressed 3-hydroxy-3-methylglutaryl-coenzyme A reductase activities in the ileum to one half the control value in the experiment where dietary fat was lard. The highly polymeric structure does not appear to be a prerequisite for nonabsorbable carbohydrate to alter lipid metabolism at least in rats fed high-fat diets.

Analysis of Variance↗

A rubella epidemic in Sasebo, Japan in 1987, with various complications.

Though the rubella vaccination programme for adolescent girls was introduced in Japan in 1977, rubella epidemics have occurred repeatedly. Also in Sasebo, Japan in 1987, we experienced various complications as follows: encephalitis (five cases), meningitis (three), thrombocytopenic purpura (four), vascular purpura (four), hemolytic anemia (two), pneumonia (eight), protein-losing gastroenteropathy (one), multiple organ disorder with encephalitis, purpura, myocarditis, hepatic and renal dysfunction (one), and congenital rubella syndrome (CRS: three). Disorders ranging over multiple organs seem to occur in acquired as well as congenital rubella infection. The incidence of encephalitis was estimated to be 1:1600 cases of rubella and two of five cases were apparently serious. Though the strategy for preventing rubella has been directed only against CRS, we should note the various and severe complications with acquired rubella infection, and should adopt two-stepped protection: vaccination of young children of both sexes and of adolescent girls.

Adolescent↗

Pernicious anemia in a patient with hypogammaglobulinemia.

A 19-year-old male with pernicious anemia and hypogammaglobulinemia (common variable immunodeficiency: CVID) is reported in comparison with classical pernicious anemia. This case was characterized by an earlier onset of anemia, the absence of autoantibodies to intrinsic factor or gastric parietal cells and involvement of the pyloric antrum as well as the gastric corpus. It is suggested that dysregulation of cellular immunity produces the autoimmune lesion in the gastric mucosa, including the pyloric antrum, in a patient with CVID, and that some of such cases develop pernicious anemia.

Adult↗

Human malignant melanoma cell line (HMV-II) for isolation of influenza C and parainfluenza viruses.

HMV-II, a human malignant melanoma cell line, was compared with other cell lines (MDCK, Vero, and LLC-MK2) and primary cultures of monkey kidney (PMK) cells for the isolation and quantification of influenza and parainfluenza viruses. HMV-II cells were superior to MDCK and LLC-MK2 cells in quantification of the influenza C virus and were used successfully in the isolation of the virus from clinical specimens. The HMV-II cell line was also more sensitive for isolating parainfluenza viruses from clinical specimens than were Vero and PMK cells; there was, however, no significant difference in the quantification of the viruses among these cultures. As far as influenza A and B viruses were concerned, the HMV-II cell line was significantly less sensitive than MDCK cells, and no virus was isolated from clinical specimens with HMV-II cells. Thus, HMV-II cells are useful for the isolation of influenza C and parainfluenza viruses as an alternative to embryonated hen's eggs and PMK cells.

Animals↗