De novo synthesis of N and P proteins as a key step in Sendai virus gene expression.
Among the members of the paramyxovirus family, the transcription process and the components involved have been studied under in vitro conditions thus far. Here, we reexamined the function of the viral RNA-dependent RNA polymerase through infection studies with Sendai virus (SeV) N and P deletion (Delta) mutants. To elucidate solely transcription-specific processes, all virus mutants also were rendered deficient in genome replication. Using mutant SeV DeltaP, the earlier suspected supplemental role of P protein was clearly demonstrated to be essential during viral gene expression. Moreover, when SeV DeltaN or DeltaN PDelta2-77 (with the 5' end of the P gene deleted) mutant was used for infections, a completely unexpected new and essential role for N protein was discovered for viral gene expression. In the early phases of an infection and in the absence of de novo viral protein synthesis, primary transcription occurs at hardly detectable levels. In contrast, if newly synthesized N protein is present, primary viral transcription reaches normal levels. From our data, we conclude that de novo synthesis of SeV N and P proteins is a key step for viral gene expression that facilitates the transition from preliminary to normal primary transcriptional activity.
Wiegand
Marian A
MA
Department of Molecular Virology, Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
Bossow
Sascha
S
Schlecht
Sabine
S
Neubert
Wolfgang J
WJ
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D019590:Nucleocapsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic; D029082:Sendai virus / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D014158:Transcription, Genetic; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D019590:Nucleocapsid Proteins; C055859:P protein, Sendai virus; D010750:Phosphoproteins; D012367:RNA, Viral; D014764:Viral Proteins