Carboxyl-terminal region of Sendai virus P protein is required for binding to viral nucleocapsids.
The Sendai virus P protein is a component of the viral nucleocapsid, where it participates in RNA synthesis. To identify domains of the protein involved in nucleocapsid recognition, deleted P protein molecules were generated from a cDNA clone of its gene. In vitro transcription of the complete gene and translation of the transcript generated a protein with electrophoretic mobility and immunoreactivity indistinguishable from those of authentic P protein. The in vitro product bound specifically to nucleocapsids when mixed with extracts from infected cells. However, a product lacking only 30 carboxyl-terminal amino acid residues (5% of the molecule) did not bind. Residues within a 195 amino acid region, adjacent to and overlapping by one amino acid with the carboxyl-terminal 30 residues, were also required for binding. No other protein region was required. Therefore, the 224-residue region which includes the carboxyl terminus appears to contain the nucleocapsid attachment site, and the 30 terminal residues either form part of the site or are required to maintain an active conformation.
Ryan
K W
KW
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Kingsbury
D W
DW
D000818:Animals; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral / Q000235:genetics; D005786:Gene Expression Regulation; D015151:Immunoblotting; D010222:Parainfluenza Virus 1, Human / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis; D014158:Transcription, Genetic; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D004279:DNA, Viral; C055859:P protein, Sendai virus; D010750:Phosphoproteins; D012367:RNA, Viral; D014758:Viral Core Proteins; D014764:Viral Proteins