Cloning of minute virus of mice cDNAs and preliminary analysis of individual viral proteins expressed in murine cells

A portion of a cDNA clone containing coding sequences for both structural proteins (VP1 and VP2) of Aleutian mink disease parvovirus (ADV) was inserted into recombinant vaccinia viruses, W:ADSP. Immunohistochemical staining of W:ADSP-infected cells revealed that the ADV antigen was readily detected and localized in the nuclei of infected cells. Analysis of W:ADSP-infected cell lysates indicated that both VP1 and VP2 were produced and comigrated with authentic VP1 and VP2 from ADV-infected Crandell feline kidney cells. These results suggested, therefore, that both VP1 and VP2 were synthesized from a single cloned transcript. CsCl density gradient centrifugation of partially purified W:ADSP-infected cell lysates indicated that the majority of the antigen was located in a fraction with a density near 1.33 g/ml, indicative of empty ADV particles. Subsequent electron microscopic examination revealed the presence of 27-nm icosahedral virion-like structures at the same density, suggesting that the proteins self-assembled into empty virions. Furthermore, sera from eight of eight mice inoculated with W:ADSP contained ADV-specific antibodies and two of these

Section: Discussioncontrasting

confidence: 82%

Expression of Aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus: self-assembly of capsid proteins into particles

Clemens

Wolfinbarger

Mori

et al. 1992

“…Capsid proteins from all four of these mutants were immunoprecipitated as previously described (31) following DNA transfection of Cosl (23), NB324K (57), and murine A92L (34) cells by the calcium phosphate precipitation method (31). As expected (13), only VP2 was detected in pMVM(D1-A1)-and pMVM (VP12635)-transfected cells and only VP1 was detected with pMVM(D2-A2) (data not shown).…”

Section: Materuils and Methodsmentioning

confidence: 57%

“…The viral capsid proteins VP1 and VP2 are translated from the viral R3 mRNAs generated from a promoter at map unit 38 (2, 31). The ratio of VPl to VP2 found in both infected cells and assembled capsids depends on which splice donor site is used to excise the small intron common to all MVM transcripts (13,29,31,42; see Fig. 1A).…”

mentioning

confidence: 99%

The minor capsid protein VP1 of the autonomous parvovirus minute virus of mice is dispensable for encapsidation of progeny single-stranded DNA but is required for infectivity

Tullis

Burger

Pintel

1993

Self Cite

The two capsid proteins of minute virus of mice, VP1 and VP2, are generated from a single large open reading frame by alternate splicing of the capsid gene mRNA. Examination of the replication of a series of mutants that express only VP1, only VP2, or neither capsid protein demonstrates that VP2 is necessary for the accumulation and encapsidation of virus progeny single-stranded DNA. VP1 is dispensable for these functions but is required to produce an infectious virion. Virus that lacks VP1 binds to cells as efficiently as wild-type minute virus of mice but fails to initiate a productive infection. Because neither capsid protein is required for viral-DNA replication, these results suggest that virus lacking VP1 is blocked at a step during virus entry, subsequent to cell binding and prior to the initiation of DNA replication.

“…2a). In this regard, expression of the minute virus of mice VP protein has been shown to be toxic to murine cells (14) and perhaps the intranuclear VP shells somehow contribute to parvoviral cytotoxicity. Alternatively, topographical segregation of replicating viral DNA and NS proteins from the VP shells might also regulate interactions between replicative intermediates.…”

Section: Panel a [Ns] With Panel B [Dna]mentioning

confidence: 99%

Subcellular localization of Aleutian mink disease parvovirus proteins and DNA during permissive infection of Crandell feline kidney cells

Oleksiewicz¹,

Costello²,

Huhtanen³

et al. 1996

Confocal microscopy allowed us to localize viral nonstructural (NS) and capsid (VP) proteins and DNA simultaneously in cells permissively infected with Aleutian mink disease parvovirus (ADV). Early after infection, NS proteins colocalized with viral DNA to form intranuclear inclusions, whereas VP proteins formedhollow intranuclear shells around the inclusions. Later, nuclei had irregular outlines and were virtually free of ADV products. In these cells, inclusions of viral DNA with or without associated NS protein were embedded in cytoplasmic VP protein. These findings implied that ADV replication within an infected cell is regulated spatially as well as temporally.