Replication of Measles Virus: Distinct Species of Short Nucleocapsids in Cytoplasmic Extracts of Infected Cells

Kiley, Michael P.; Gray, Robert H.; Payne, Francis E.

doi:10.1128/jvi.13.3.721-728.1974

Cited by 27 publications

(21 citation statements)

References 18 publications

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“…Besides being present in the intact (infectious) virion fraction, measles polymerase activity was also associated with measles nucleocapsids, as was also noted for other paramyxoviruses (11,21), since it was found associated with the IP-3 isolate and was found sedimenting at a velocity and a density consistent with those reported for measles nucleocapsids (13).…”

Section: Discussionsupporting

confidence: 73%

“…Figure 1B shows that the RNA polymerase activity shifted to the density region of nucleocapsids. Nucleocapsids prepared in this manner had associated polymerase activity, which sedimented at 200S in a sucrose velocity gradient (data not shown) as reported for measles nucleocapsids (13). The activity was associated with a positive complement fixation test for measles nucleocapsids.…”

Section: Ment-fixing Antigen and May Contain Measlessupporting

confidence: 64%

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Characterization of an RNA-dependent RNA polymerase activity associated with measles virus

Seifried¹,

Albrecht²,

Milstien³

1978

J Virol

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An RNA-dependent RNA polymerase activity has been found copurifying with measles virus infectivity and complement-fixing antigen in three Vero cell-grown variants of measles virus: the attenuated Edmonston B strain, the natural nonattenuated Edmonston strain, and a subacute sclerosing panencephalitis isolate, IP-3. Incubation of purified measles virions with immunoglobulin G derived from sera of monkeys hyperimmunized against measles specifically removes activity sedimenting in the density region of measles virions. The requirements of the reaction, which is RNase sensitive, are similar to those reported for other paramyxovirus-associated activities, including detergent, divalent cation, ribonucleoside triphosphates, and a reducing agent. The size classes of RNA synthesized correspond to those found in measles-infected cells, including 50, 35, and 16 to 20S. The product RNA of the Edmonston B virus-stimulated reaction was rendered RNase resistant by annealing with RNA extracted from purified Edmonston B virions. RNA from uninfected Vero cells was ineffective in the annealing reaction. Measles virus has been classified as a paramyxovirus on the basis of morphology and RNA size (8, 19, 23, 24). Reports in the literature of RNA-dependent RNA polymerase in the virions of other paramyxoviruses (4, 11, 18, 22) suggested that measles virus might contain a similar activity. Here we describe and characterize a measles virus-associated RNA-dependent RNA polymerase activity. MATERIALS AND METHODS Cells and viruses. Vero cells, a continuous line of African green monkey kidney cells, were supplied by the Cell Biology Section, Bureau of Biologics, and were used both for virus production and infectivity assays (2). They were grown and maintained in Eagle minimal essential medium supplemented with 10% fetal calf serum, 0.03% glutamine, 50 ,ug of gentamicin per ml, and 0.4 pLg of amphotericin B per ml.

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Section: Discussionsupporting

confidence: 73%

Section: Ment-fixing Antigen and May Contain Measlessupporting

confidence: 64%

Characterization of an RNA-dependent RNA polymerase activity associated with measles virus

Seifried¹,

Albrecht²,

Milstien³

1978

J Virol

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“…The pattern of RNA synthesis in measles infected cells has also been described [7,12]. In addition, we have recently reported on the presence of several distinct size classes of virus specific nueleocapsids in measles virus infected cells [6]. The largest nucleocapsid structure contains the putative complete viral RNA (i. e. 50S) while the smallest structure contains RNA which is approximately one-tenth the size of 50S RNA.…”

Section: Introductionmentioning

confidence: 66%

“…Extraction of RNA from sucrose gradient fractions has been described previously [8]. Analysis of this RNA in 5--20~ sucrose gradients prepared in 0.1 M acetate buffer (pH 5.0) has also been described [6].…”

Section: Labeling and Analysis Of Infected Cell Extracts Vero Cell Mmentioning

confidence: 99%

“…1 shows the presence of two major virus specific labeled structures with sedimentation coefficients of approximately 200S and ll0S. The RNA associated with the 200S nucleocapsids is nearly 100~ resistant to RNAase and that from the ll0S structures is also completely resistant when polyribosomes are removed from that region of the gradient by pretreatment with EDTA [6].…”

Section: Characterization O~ Yirusspeci]ic Nucleocapsid~ From Mea~lesmentioning

confidence: 99%

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Evidence of precursors of defective measles virus

Kiley

Payne

1974

Med Microbiol Immunol

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Cytoplasmic extracts of Vero cells infected with wild strain Edmonston measles virus were found to contain at least two distinct nueleocapsid species. The two most prominent species of nucleocapsids sedimented at 200S and ll0S and contained RNA of molecular weight 6.0 • 10 e and 0.6 • l0 s daltons respectively. Both species of nucleocapsids had a density of 1.31 g/cm 3 in CsC1. A third species sedimenting at 170S was not present in all experiments and was not characterized in detail. Infection of cells with undiluted-passage virus usually resulted in production of mostly ll0S nucleoeapsids while both ll0S and 200S species were found when diluted-passage virus was used. These results suggest that measles virus may produce distinct classes of defective virus which contain segments of RNA representing as little as 10% of the complete viral genome.

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Immunogenetic Analysis and Serum Viral Antibody Titers in Multiple Sclerosis

Jn¹,

Kl²,

Gn³

et al. 1976

Archives of Neurology

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In order to investigate the possible relationships between multiple sclerosis (MS) and the factors of genetic predisposition and exposure to infectious agents, studies were undertaken in 59 male patients with MS to determine the histocompatibility antigen (HL-A) type and the serum antibody titer to rubeola hemagglutinin (HA), rubeola envelope antigen (V), rubeola nucleocapsid antigen (S), rubella, herpes simplex, cytomegalovirus, and parainfluenza 1. The incidence of HL-A7 was significantly higher, and HL-A12 incidence was significantly lower, in the MS group than in control groups. The geometric mean titers to rubeola HA and to the V and S antigens were lower in the HL-A7 positive control patients than in either the non-HL-A7 controls or the MS patients. There were no specific relationships of the rubeola titer to HL-A12. Antibody titers to the other viruses studied were not significantly different in MS patients as compared to controls, nor was there any relationship of HL-A7 or HL-A12 and serum titer to those viruses. This study provides further evidence of a genetic predisposition to MS, and of a relationship between rubeola and this disease. These findings suggests that the failure of MS to appear in individuals who might be genetically predisposed to this disorder may be related to their immune response to rubeola.

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Replication of Measles Virus: Distinct Species of Short Nucleocapsids in Cytoplasmic Extracts of Infected Cells

Cited by 27 publications

References 18 publications

Characterization of an RNA-dependent RNA polymerase activity associated with measles virus

Characterization of an RNA-dependent RNA polymerase activity associated with measles virus

Evidence of precursors of defective measles virus

Immunogenetic Analysis and Serum Viral Antibody Titers in Multiple Sclerosis

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