Replication of minute virus of mice DNA is critically dependent on accumulated levels of NS2.
Following transfection of murine fibroblasts, the lymphotropic strain of minute virus of mice (MVMi) does not efficiently produce progeny single-strand DNA (ssDNA). However, changing a single nucleotide in the MVMi 3' splice site to that found in the fibrotropic strain MVMp enabled full DNA replication and production of ssDNA. This change enhanced excision of the large intron and the production of NS2, likely by improving interaction, in fibroblasts with the branch point-binding U2 snRNA. One function of NS2 involves interaction with the nuclear export protein Crm1. The defect in production of MVMi ssDNA in fibroblasts can also be overcome by introducing a mutation in MVMi NS2 that enhances its interaction with Crm1. Although MVMi contains a 3' splice site that performs poorly in fibroblasts, MVMi generated at least as much R2 and NS2 in murine lymphocytes as did MVMp in fibroblasts. Therefore, it appears that MVMp has acquired a mutation that improves the excision of the large intron, as it adapted to fibroblasts to accommodate the need for NS2 for replication in these cells, and that the ratio of NS1 to NS2 may play a larger role in the host range of MVM than previously appreciated.
Choi
Eun-Young
EY
Department of Molecular Microbiology and Immunology, Life Sciences Center, School of Medicine, University of Missouri-Columbia, 65211-7310, USA.
Newman
Ann E
AE
Burger
Lisa
L
Pintel
David
D
D000818:Animals; D002460:Cell Line; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000096:biosynthesis*; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D005347:Fibroblasts / Q000821:virology; D007438:Introns / Q000235:genetics; D051379:Mice; D008915:Minute Virus of Mice / Q000235:genetics* / Q000502:physiology*; D009154:Mutation; D022821:RNA Splice Sites / Q000235:genetics; D012342:RNA, Small Nuclear / Q000378:metabolism; D012367:RNA, Viral / Q000032:analysis / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D022821:RNA Splice Sites; D012342:RNA, Small Nuclear; D012367:RNA, Viral; C405484:U2 small nuclear RNA; D017361:Viral Nonstructural Proteins