Two separate domains within vesicular stomatitis virus phosphoprotein support transcription when added in trans.
The structural phosphoprotein NS of vesicular stomatitis virus, in association with the virion-associated RNA polymerase L protein, transcribes the genome ribonucleoprotein template in vitro. It contains an acidic N-terminal domain and two distinct domains at the C-terminal end that are involved in binding to the polymerase protein and the template RNA enwrapped with the nucleocapsid protein. In the present study, the portions of the NS gene that encode the N- and C-terminal domains of the protein were cloned in pGEM vectors and expressed by in vitro transcription and translation. It was shown that two polypeptides obtained by translation of the encoded mRNAs support RNA synthesis in vitro in a reconstitution reaction when they are added together in trans. Moreover, the N-terminal domain can be functionally substituted by structurally similar polypeptides.
Chattopadhyay
D
D
Roche Institute of Molecular Biology, Roche Research Center, Nutley, NJ 07110.
Banerjee
A K
AK
D002213:Capsid / Q000502:physiology*; D003001:Cloning, Molecular; D007526:Isoelectric Point; D010750:Phosphoproteins / Q000502:physiology*; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000502:physiology; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*; D014758:Viral Core Proteins / Q000502:physiology*; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000502:physiology
D010750:Phosphoproteins; D011994:Recombinant Proteins; D014157:Transcription Factors; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins