Molecular cloning and expression of human parainfluenza virus type 1 L gene.
The large (L) protein, a subunit of paramyxovirus RNA polymerase complex is responsible for the majority of enzymic activities involved in viral replication and transcription. To gain insight of the functions of the L protein, we cloned the L gene of human parainfluenza virus type 1 (hPIV1) and sequenced the entire gene. The L gene, which was 6800 nucleotides, encoded a protein of 2223 residues with a calculated molecular weight of 253657. The predicted amino acid sequence was highly homologous with that of Sendai virus (SV) L (86% identity). The hPIV1 L protein expressed from the cloned L gene bound hPIV1 P expressed in the same cells. When cells were transfected with hPIV1 L, P and NP genes together with SV minigenome RNA containing a chloramphenicol acetyltransferase (CAT) gene (Send-CAT), RNA was transcribed, and CAT proteins were detected. These results indicate that the protein encoded by the cloned hPIV1 L gene was biologically functional and that the hPIV1 polymerase complex recognized SV transcription initiation and termination sequences to produce viral transcripts.
Takimoto
T
T
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, 332 N. Lauderdale, Memphis, TN 38105, USA. toru.takimoto@stjude.org
Bousse
T
T
Portner
A
A
D000595:Amino Acid Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000378:metabolism; D010222:Parainfluenza Virus 1, Human / Q000235:genetics* / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D016415:Sequence Alignment; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D019590:Nucleocapsid Proteins; D010750:Phosphoproteins; D012367:RNA, Viral; D014764:Viral Proteins; C068855:P protein, parainfluenza virus 1; D015500:Chloramphenicol O-Acetyltransferase; D012321:DNA-Directed RNA Polymerases