Transcriptase activity associated with rabies virion

Kawai, Akihiko

doi:10.1128/jvi.24.3.826-835.1977

Cited by 73 publications

(38 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The p35 was usually detected in a more acidic region of the 2-D gel than was p37-0/p37-1, which may be due to truncation of some basic amino acid-rich region from the N-terminus (6,31). In addition, the pI value of p35 was not altered by treatment with the phosphatase, implying that p35 is not a target of additional phosphorylation or dephosphorylation after the synthesis, and might not be involved functionally in the virus replication process, although a small amount of p35 is also detected in the virion (16,31). From these considerations, we assume that the N-terminal region of the P protein is essential for the N-P complex formation and the hyperphosphorylation.…”

Section: Discussionmentioning

confidence: 95%

“…The p40 was detected mostly as an NC-associated form in the cell and in the virion (16,31), however, its role(s) and function(s) still remained obscure. Relative content of p40 was not so abundant in the infected cells, but treatment of the infected cells with okadaic acid induced accumulation of p40 (31), and also resulted in production of more acidic forms of p40 (e.g., p40-2, p40-3, p40-4).…”

Section: Discussionmentioning

confidence: 99%

“…Virus purification. Rabies virions propagated in BHK-21 cell monolayer cultures were recovered and concentrated from the culture medium by precipitation with polyethylene glycol and purified by sucrose density gradient centrifugation as described previously (16). Virions recovered from a sharp band formed in the gradient were concentrated by centrifugation again at 68,000 g for 1.5 hr.…”

Section: Methodsmentioning

confidence: 99%

“…From the analogy of the VSV P protein, the rabies virus P protein is also thought to serve similarly as a molecular chaperone and a non-catalytic subunit of the viral RNA polymerase (18). As to the heterogeneity of the rabies virus P protein, we reported previously that, in addition to a major form (37 kDa) of the P protein, the virion contained another minor, but full-sized, component which was recognized by anti-P antibody and termed p40 according to its relative electrophoretic mobility in SDS-PAGE (16,31). The p40 was very little in amount in the cell, and was more phosphorylated than a major 37-kDa component (p37) of the P protein (6,20,32).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Studies on the Rabies Virus RNA Polymerase: 3. Two‐Dimensional Electrophoretic Analysis of the Multiplicity of Non‐Catalytic Subunit (P Protein)

Eriguchi

Toriumi

Kawai

2002

Microbiology and Immunology

View full text Add to dashboard Cite

The genome of rabies virus encodes two kinds of phosphoproteins; one is the nucleoprotein (N) which encapsidates viral genomic RNA, and the other is a nominal phosphoprotein (P; formerly called M1), that is thought to work multifunctionally for viral RNA synthesis. Newly synthesized N proteins are mostly associated with P proteins, which is thought to be a prerequisite step for the N protein to be used for encapsidating viral genomic and anti-genomic RNAs specifically. Phosphorylation of the N protein occurs during or after the encapsidation (20). The P protein also works for viral RNA synthesis with the viral large (L) protein (catalytic subunit of the viral RNA polymerase), but the role(s) of the P protein and the mechanism of its phosphorylation remained to be elucidated precisely (18,33).The P protein of another rhabdovirus family (vesicular stomatitis virus; VSV) has been studied extensively (see a review by Banerjee and Barik; 1). The P protein was shown to serve as a molecular chaperone for the viral 32 P]phosphate implied that p40 was a hyperphosphorylated form. We further examined here these proteins by two-dimensional (2-D) gel electrophoresis and immunoblotting, showing that a major component, p37, was composed of multiply modified subcomponents of different pIs (termed p37-1, p37-2, p37-3, etc., based on their acidity) in the virion and infected cells, but the unmodified precursor (termed p37-0) was little in amount. The viral nucleocapsid (NC)-bound P proteins were composed of multiple forms of p37 (the major one was p37-1) and also a minor component, p40-1. P proteins which were bound to newly synthesized free N proteins were mostly composed of p37-1, indicating that hyperphosphorylation of P proteins occurred after their being used for the encapsidation. Treatment of the infected cells with okadaic acid induced accumulation of the more acidic forms of P proteins, suggesting that heterogeneity in the full-sized P proteins is a reflection of their dynamic aspects of multiple cycles of phosphorylations and dephosphorylations in the cell. Two-D gel analyses demonstrated also that p40 was not so acidic as we expected, and implied that our previous data of apparent hyperphosphorylation of p40 was due to very frequently recycled utilization of the protein, and preformed non-labeled P proteins were also 32 P-phosphorylated in a radiolabeling period and were converted to the p40.Key words: rabies virus P protein, multiple components of P protein, non-catalytic subunit of rabies virus RNA polymerase, okadaic acid, phosphorylation and dephosphorylation, isoelectric focusing, 2-D gel electrophoresis

show abstract

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Studies on the Rabies Virus RNA Polymerase: 3. Two‐Dimensional Electrophoretic Analysis of the Multiplicity of Non‐Catalytic Subunit (P Protein)

Eriguchi

Toriumi

Kawai

2002

Microbiology and Immunology

View full text Add to dashboard Cite

show abstract

“…In a tentative list of host-derived virion components, we may be able to identify the cellular factors that are deeply involved in the virus replication process. From this viewpoint, we have been involved in identifying the host-derived minor components in enveloped virions since we noticed many years ago that the highly purified rabies virion preparations still contained several kinds of host-derived minor but regular components (9). One such component was identified as the actin by our colleagues (17).…”

mentioning

confidence: 99%

The Rabies Virion‐Associated 100‐kDa Polypeptide (VAP100) Is a Host‐Derived Minor Component of the Viral Envelope

Xiao

Komiya

Tochikura

et al. 2000

Microbiology and Immunology

View full text Add to dashboard Cite

We investigated a minor polypeptide component of 100-kDa detected in the rabies virion (referred to as VAP100) by using a monoclonal antibody (mAb), #16743, which was shown to recognize the SDS-denatured VAP100 antigen by immunoblot analyses. Although the VAP100 antigen was hardly detectable in the cell by usual immunoblot methods with this mAb, we could detect the antigen by a luminescent immunoblot method as well as by immunoprecipitation from the metabolically radiolabeled cell lysates and virions. Fluorescent antibody (FA) staining with mAb #16743 detected the uniformly distributed antigen on the formalin-fixed normal BHK-21 cells, while slight accumulation of the antigen was also seen in the Golgi area when the cells were permeabilized by treatment with Triton X-100 after fixation. Rabies virus infection induced alteration of the behavior of VAP100 to show a spotted distribution pattern in virusinfected cells. Double FA staining with mAb #16743 and rabbit antibody against the rabies virus envelope antigen demonstrated colocalized distribution of the viral envelope antigens and VAP100 in the cell. From these results, we think that VAP100 is a membrane-associated component of the cell, and its colocalized distribution with the viral envelope antigens in the cell implicates an intimate association of the VAP100 with viral envelope protein(s) and a reflection of possible involvement in the efficient incorporation of VAP100 into the virion.

show abstract

Isolation and characterization of tunicamycin resistant mutants from Chinese Hamster ovary cells

Sudo

Onodera

1979

Journal Cellular Physiology

View full text Add to dashboard Cite

Stable clones selected for resistance to tunicamycin (TM) have been isolated from Chinese Hamster Ovary (CHO) cells. The TMR phenotype is stable for more than nine months in the absence of the drug. The morphology of TMR mutant varies from epitheloid to abnormally elongate. The mutants do not display cross-resistance for ConA but are slightly cross-resistant to PHA. Biochemically labeled membrane proteins and glycoprotein of Vesicular stomatitis virus (VSV) grown in the TMR mutants revealed that the incorporation of radioactive glucosamine was markedly reduced in the mutants. The results indicate that TMR cells are a novel type of membrane mutant.

show abstract

Transcriptase activity associated with rabies virion

Cited by 73 publications

References 24 publications

Studies on the Rabies Virus RNA Polymerase: 3. Two‐Dimensional Electrophoretic Analysis of the Multiplicity of Non‐Catalytic Subunit (P Protein)

Studies on the Rabies Virus RNA Polymerase: 3. Two‐Dimensional Electrophoretic Analysis of the Multiplicity of Non‐Catalytic Subunit (P Protein)

The Rabies Virion‐Associated 100‐kDa Polypeptide (VAP100) Is a Host‐Derived Minor Component of the Viral Envelope

Isolation and characterization of tunicamycin resistant mutants from Chinese Hamster ovary cells

Contact Info

Product

Resources

About