Human parainfluenza virus type 3 transcription in vitro: role of cellular actin in mRNA synthesis.
Purified ribonucleoprotein complexes of human parainfluenza virus type 3 (HPIV-3) virions required, in addition to the viral proteins, soluble cytoplasmic proteins from uninfected cells for the synthesis of mRNAs in vitro. In contrast to Sendai virus transcription, in vitro RNA synthesis from HPIV-3 ribonucleoprotein complexes was not stimulated significantly by purified tubulin. Moreover, cytoplasmic extract depleted of tubulin by immunoprecipitation stimulated HPIV-3 transcription effectively, suggesting involvement of a host protein(s) other than tubulin in the HPIV-3 transcription process. The transcription stimulatory factor was purified from uninfected cell extract by conventional chromatography and was found to contain a major 43-kDa polypeptide. In Western blot (immunoblot) analysis, this protein reacted with antiactin antibody, suggesting that the 43-kDa polypeptide is actin. This possibility was further supported by its polymerization activity and properties of binding to blue-Sepharose and heparin-Sepharose columns. Furthermore, when the cell extract was depleted of actin by immunoprecipitation by antiactin antibody, the stimulatory activity was abolished, indicating an involvement of actin in the stimulation of HPIV-3 transcription. After purification from RNAses, similar stimulatory activity associated with the 43-kDa protein was detected in other cell lines as well, including CV-1, HeLa, and BHK.
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Department of Molecular Biology, Cleveland Clinic Foundation, Ohio 44195-5069.
Lesoon
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Banerjee
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D000199:Actins / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D008970:Molecular Weight; D010222:Parainfluenza Virus 1, Human / Q000235:genetics; D010224:Parainfluenza Virus 3, Human / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis*; D012261:Ribonucleoproteins / Q000235:genetics*; D014158:Transcription, Genetic; D014404:Tubulin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics*; D014771:Virion / Q000235:genetics
D000199:Actins; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D014404:Tubulin; D014764:Viral Proteins