A replicative form of the DNA of minute virus of mice

1973 ◽  
Vol 19 (1) ◽  
pp. 35-41 ◽  
Author(s):  
Phyllis R. Dobson ◽  
C. W. Helleiner
Keyword(s):  
Sodium Dodecylsulfate ◽  
Plaque Assay ◽  
Progeny Virus ◽  
Sedimentation Rates ◽  
Free Virus ◽  
Minute Virus Of Mice ◽  
Selective Precipitation ◽  
L Cells ◽  
Minute Virus ◽  
Cellular Dna

The growth of minute virus of mice (MVM) in L cells was followed by plaque assay of both cell-associated and free virus at intervals up to 36 h after infection. The major production of progeny virus occurred after incubation for 27 h and most of the virus remained cell-associated. L cells infected with MVM were pulse-labeled with 3H-thymidine for 6 h preceding induction of lysis with sodium dodecylsulfate. Two new species of DNA remained in the supernatant fractions from lysates after selective precipitation by 1 M NaCl of cellular DNA. These first appeared 17 h after infection. On the basis of sedimentation rates and response to heating and to treatment with alkali, one of these species is a hydrogen-bonded duplex molecule, which on denaturation hybridized with the single-stranded DNA of MVM virions.

scholarly journals Parvovirus minute virus of mice interacts with sites of cellular DNA damage to establish and amplify its lytic infection

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Kinjal Majumder ◽  
Juexin Wang ◽  
Maria Boftsi ◽  
Matthew S Fuller ◽  
Jordan E Rede ◽  
...  
Keyword(s):  
Dna Damage ◽  
Virus Genome ◽  
Chromosome Conformation ◽  
Minute Virus Of Mice ◽  
Capture Assay ◽  
Genome Wide ◽  
Topologically Associated Domains ◽  
Infected Cells ◽  
Minute Virus ◽  
Cellular Dna

We have developed a generally adaptable, novel high-throughput Viral Chromosome Conformation Capture assay (V3C-seq) for use in trans that allows genome-wide identification of the direct interactions of a lytic virus genome with distinct regions of the cellular chromosome. Upon infection, we found that the parvovirus Minute Virus of Mice (MVM) genome initially associated with sites of cellular DNA damage that in mock-infected cells also exhibited DNA damage as cells progressed through S-phase. As infection proceeded, new DNA damage sites were induced, and virus subsequently also associated with these. Sites of association identified biochemically were confirmed microscopically and MVM could be targeted specifically to artificially induced sites of DNA damage. Thus, MVM established replication at cellular DNA damage sites, which provide replication and expression machinery, and as cellular DNA damage accrued, virus spread additionally to newly damaged sites to amplify infection. MVM-associated sites overlap significantly with previously identified topologically-associated domains (TADs).

scholarly journals Regulation of Minute Virus of Mice NS1 Replicative Functions by Atypical PKCλ In Vivo

2003 ◽  
Vol 77 (1) ◽  
pp. 433-442 ◽  
Author(s):  
Jürg P. F. Nüesch ◽  
Sylvie Lachmann ◽  
Romuald Corbau ◽  
Jean Rommelaere
Keyword(s):  
Dna Replication ◽  
Dominant Negative ◽  
Progeny Virus ◽  
Ns1 Protein ◽  
Viral Dna ◽  
Minute Virus Of Mice ◽  
Viral Dna Replication ◽  
Minute Virus

ABSTRACT Minute virus of mice NS1 protein is a multifunctional phosphoprotein endowed with a variety of enzymatic and regulatory activities necessary for progeny virus particle production. To regulate all of its different functions in the course of a viral infection, NS1 has been proposed to be modulated by posttranslational modifications, in particular, phosphorylation. Indeed, it was shown that the NS1 phosphorylation pattern is altered during the infectious cycle and that the biochemical profile of the protein is dependent on the phosphorylation state of the polypeptide. Moreover, in vitro approaches have identified members of the protein kinase C (PKC) family, in particular, atypical PKC, as regulators of viral DNA replication through the phosphorylation of NS1 residues T435 and S473, thereby activating the protein for DNA unwinding activities. In order to substantiate these findings in vivo, we produced NS1 in the presence of a dominant-negative PKCλ mutant and characterized the purified protein in vitro. The NS1 protein produced under these conditions was found to be only partially phosphorylated and as a consequence to be deficient for viral DNA replication. However, it could be rescued for this viral function by treatment with recombinant activated PKCλ. Our data clearly demonstrate that NS1 is a target for PKCλ phosphorylation in vivo and that this modification is essential for the helicase activity of the viral polypeptide. In addition, the phosphorylation of NS1 at residues T435 and S473 appeared to occur mainly in the nucleus, providing further evidence for the involvement of PKCλ which, unlike PKCζ, accumulates in the nuclear compartment of infected cells.

scholarly journals Biochemical Activities of Minute Virus of Mice Nonstructural Protein NS1 Are Modulated In Vitro by the Phosphorylation State of the Polypeptide

1998 ◽  
Vol 72 (10) ◽  
pp. 8002-8012 ◽  
Author(s):  
Jürg P. F. Nüesch ◽  
Romuald Corbau ◽  
Peter Tattersall ◽  
Jean Rommelaere
Keyword(s):  
Posttranslational Modifications ◽  
Nonstructural Protein ◽  
Virus Production ◽  
Progeny Virus ◽  
Helicase Activity ◽  
Minute Virus Of Mice ◽  
Target Dna ◽  
Minute Virus ◽  
Cytotoxic Stress

ABSTRACT NS1, the 83-kDa major nonstructural protein of minute virus of mice (MVM), is a multifunctional nuclear phosphoprotein which is required in a variety of steps during progeny virus production, early as well as late during infection. NS1 is the initiator protein for viral DNA replication. It binds specifically to target DNA motifs; has site-specific single-strand nickase, intrinsic ATPase, and helicase activities; trans regulates viral and cellular promoters; and exerts cytotoxic stress on the host cell. To investigate whether these multiple activities of NS1 depend on posttranslational modifications, in particular phosphorylation, we expressed His-tagged NS1 in HeLa cells by using recombinant vaccinia viruses, dephosphorylated it at serine and threonine residues with calf intestine alkaline phosphatase, and compared the biochemical activities of the purified un(der)phosphorylated (NS1O) and the native (NS1P) polypeptides. Biochemical analyses of replicative functions of NS1O revealed a severe reduction of intrinsic helicase activity and, to a minor extent, of ATPase and nickase activities, whereas its affinity for the target DNA sequence [ACCA]2–3 was enhanced compared to that of NS1P. In the presence of endogenous protein kinases found in replication extracts, NS1O showed all functions necessary for resolution and replication of the 3′ dimer bridge, indicating reactivation of NS1O by rephosphorylation. Partial reactivation of the helicase activity was found as well when NS1O was incubated with protein kinase C.

scholarly journals Author response: Parvovirus minute virus of mice interacts with sites of cellular DNA damage to establish and amplify its lytic infection

2018 ◽  
Author(s):  
Kinjal Majumder ◽  
Juexin Wang ◽  
Maria Boftsi ◽  
Matthew S Fuller ◽  
Jordan E Rede ◽  
...  
Keyword(s):  
Dna Damage ◽  
Lytic Infection ◽  
Author Response ◽  
Minute Virus Of Mice ◽  
Minute Virus ◽  
Cellular Dna

scholarly journals Interrelation between Viral and Cellular DNA Synthesis in Mouse Cells Infected with the Parvovirus Minute Virus of Mice

1983 ◽  
Vol 64 (9) ◽  
pp. 1991-1998 ◽  
Author(s):  
N. Hardt ◽  
C. Dinsart ◽  
S. Spadari ◽  
G. Pedrali-Noy ◽  
J. Rommelaere
Keyword(s):  
Dna Synthesis ◽  
Minute Virus Of Mice ◽  
Minute Virus ◽  
Mouse Cells ◽  
Cellular Dna
Sign in / Sign up

Export Citation Format

Share Document