Interaction of cellular tubulin with Sendai virus M protein regulates transcription of viral genome.
Cellular tubulin has been shown to activate in vitro transcription with Sendai virus (SeV) particles. In this study, the molecular basis for the transcriptional activation by tubulin was investigated. We showed that tubulin dissociates viral matrix (M) protein, which acts as a negative regulator for transcription, from viral ribonucleoprotein (RNP) consisting of L, P, N proteins, and the genome RNA. Both alpha and beta subunits of human tubulin, which were expressed as GST fusion proteins, were found to stimulate viral mRNA synthesis similar to native alpha/beta-heterodimer tubulin. Pull-down assay using GST-tubulin subunits demonstrated that M protein is released from the RNP as a complex with each tubulin subunit. In vitro-binding analyses revealed that M protein directly interacts with tubulin as well as microtubules. These findings suggest that interaction of M protein with tubulin may have an important role in the regulation of SeV transcription.
Department of Biochemistry, School of Pharmaceutical Sciences, Kitasato University, Shirokane, Minato-ku, Tokyo 108-8641, Japan.
Tomoaki
T
Ogino
Minako
M
Iwama
Yuki
Y
Ohsawa
Kiyohisa
K
Mizumoto
D001665:Binding Sites; D002478:Cells, Cultured; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D016679:Genome, Viral; D006801:Humans; D008870:Microtubules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D021122:Protein Subunits; D029082:Sendai virus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation / Q000502:physiology*; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000378:metabolism*
C097020:M protein, Sendai virus; D021122:Protein Subunits; D014404:Tubulin; D014763:Viral Matrix Proteins