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Sequence determination of the mumps virus HN gene
M N Waxham1, J Aronowski, A C Server
1Department of Neurology, University of Texas Health Science Center, Houston 77225.
Abstract:
The hemagglutinin-neuraminidase protein (HN) of mumps virus was purified by immunoaffinity chromatography and fragmented by the combined action of CNBr and trypsin. The resulting peptides were separated by HPLC and sequenced by automated Edman degradation. Using this HN-specific amino acid sequence data, a degenerate oligonucleotide was produced and subsequently used to screen a mumps virus cDNA library to isolate HN-specific clones. The complete nucleotide sequence of the HN gene was determined. The monocistronic HN mRNA is approximately 1900 nucleotides long and encodes a single open reading frame of 582 amino acids. The HN protein has a unique hydrophobic stretch of 19 amino acids at its N-terminus that apparently anchors the protein in the viral envelope. A comparison of the mumps virus HN protein sequence with the sequences of the other known paramyxovirus HNs indicates that mumps virus is most closely related to SV-5, followed in decreasing order by NDV, parainfluenza virus 3, and Sendai virus.
Insights
Researchers sequenced the mumps virus hemagglutinin-neuraminidase (HN) protein, revealing its complete gene sequence and evolutionary relationships within the paramyxovirus family.
Area of Science:
- Virology
- Molecular Biology
- Protein Chemistry
Background:
- The hemagglutinin-neuraminidase (HN) protein is crucial for mumps virus infectivity.
- Understanding HN's genetic and structural properties aids in viral classification and vaccine development.
Purpose of the Study:
- To determine the complete nucleotide sequence of the mumps virus HN gene.
- To analyze the deduced amino acid sequence of the HN protein.
- To compare the mumps virus HN with other paramyxovirus HN proteins.
Main Methods:
- Immunoaffinity chromatography for HN protein purification.
- Peptide fragmentation using CNBr and trypsin.
- High-performance liquid chromatography (HPLC) for peptide separation.
- Automated Edman degradation for peptide sequencing.
- cDNA library screening using degenerate oligonucleotides.
Main Results:
- The complete nucleotide sequence of the mumps virus HN gene was determined.
- The HN gene encodes a 582-amino acid protein with a unique N-terminal hydrophobic anchor.
- Mumps virus HN shows closest evolutionary relationship to SV-5 among paramyxoviruses.
Conclusions:
- The determined HN sequence provides insights into mumps virus molecular biology.
- Sequence homology suggests mumps virus is evolutionarily linked to SV-5.
- The findings contribute to the broader understanding of paramyxovirus diversity.