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Sequence characterization and expression of the matrix protein gene of human parainfluenza virus type 1
U F Power1, K W Ryan, A Portner
1Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101.
Abstract:
The nucleotide sequence of the M gene of human parainfluenza virus type 1 (hPIV1) was determined from genomic RNA and cDNA copies of the entire gene. The M gene contained 1173 nucleotides. It had one large open reading frame capable of encoding a protein of 348 amino acids (M(r) = 38,404). The predicted amino acid sequence of the hPIV1 M protein is highly basic (+20 at neutral pH). A pGEM-1 expression vector containing the M gene was used for cell-free transcription and translation. The resultant protein was confirmed to be M by electrophoretic mobility and immunoprecipitation. Among other paramyxoviridae the hPIV1 M amino acid sequence was most closely related to the Sendai virus M sequence (87% identity). The pattern of M gene relatedness observed from the alignment of 16 paramyxoviridae M protein amino acid sequences was not predicted by the viruses' taxonomic classification.
Insights
Researchers sequenced the M gene of human parainfluenza virus type 1 (hPIV1), revealing a protein highly related to Sendai virus. This finding challenges current taxonomic classifications for paramyxoviruses.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Human parainfluenza virus type 1 (hPIV1) is a significant respiratory pathogen.
- Understanding viral gene sequences is crucial for developing antiviral strategies and vaccines.
- The M gene's role and sequence variation across paramyxoviruses require further investigation.
Purpose of the Study:
- To determine the complete nucleotide sequence of the hPIV1 M gene.
- To characterize the predicted amino acid sequence and properties of the hPIV1 M protein.
- To compare the hPIV1 M protein sequence with those of other paramyxoviruses.
Main Methods:
- Genomic RNA and complementary DNA (cDNA) were used to determine the M gene sequence.
- Bioinformatic analysis predicted the open reading frame and amino acid sequence.
- Cell-free transcription and translation using an expression vector confirmed protein identity.
- Sequence alignment and phylogenetic analysis were performed for comparative studies.
Main Results:
- The hPIV1 M gene consists of 1173 nucleotides, encoding a 348-amino acid protein.
- The predicted hPIV1 M protein is highly basic.
- The hPIV1 M protein sequence exhibits 87% identity to the Sendai virus M protein.
- Sequence relatedness did not align with the established taxonomic classification of paramyxoviruses.
Conclusions:
- The M gene sequence of hPIV1 has been fully elucidated.
- The high sequence similarity between hPIV1 and Sendai virus M proteins suggests potential functional or evolutionary links.
- These findings highlight the need to re-evaluate the taxonomic relationships within the Paramyxoviridae family based on molecular data.