Early and late gene expression in UT-7 cells infected with B19 parvovirus.
UT-7, a human megakaryocytoblastoid cell line, can be persistently infected with B19 parvovirus. We performed detailed serial analysis of parvovirus DNA replication and RNA transcription of synchronized cells. RNA transcription appeared as an early event following infection, with viral RNA detected about 6 hr after infection. In contrast, dimer-replicative intermediate forms of parvovirus DNA did not appear until more than 16 hr after infection. Northern analysis of specific transcripts showed an earlier appearance of nonstructural protein RNA (6 hr) compared to capsid protein RNA (24 hr). The addition of an inhibitor of protein synthesis to block synthesis of nonstructural protein abolished capsid protein RNA transcription as well as DNA replication. Primer extension analysis confirmed the initiation of all transcription from the single P6 promoter. RNA transcription precedes DNA replication of B19 parvovirus in these cells, and RNA processing may have a major role in regulating gene expression.
Shimomura
S
S
Cell Biology Section, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.
Wong
S
S
Brown
K E
KE
Komatsu
N
N
Kajigaya
S
S
Young
N S
NS
D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis; D005814:Genes, Viral / Q000235:genetics*; D006801:Humans; D007947:Leukemia, Megakaryoblastic, Acute; D008969:Molecular Sequence Data; D016732:Parvovirus B19, Human / Q000235:genetics* / Q000254:growth & development; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D014764:Viral Proteins / Q000032:analysis; D015678:Viral Structural Proteins / Q000235:genetics
D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins; D015678:Viral Structural Proteins; D003513:Cycloheximide