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Virus-like particles and E1-E4 protein expressed from the human papillomavirus type 11 bicistronic E1-E4-L1
D R Brown1, L Pratt, J T Bryan
1Department of Medicine, Indiana University School of Medicine, Indianapolis 46202, USA.
Virology
|August 1, 1996
Summary
This study demonstrates that human papillomavirus type 11 (HPV 11) E1-E4 and L1 proteins can be expressed from a single bicistronic mRNA. This finding is crucial for understanding HPV 11 viral assembly and pathogenesis.
Area of Science:
- Virology
- Molecular Biology
- Protein Expression
Background:
- Detection of human papillomavirus type 11 (HPV 11) E1-E4 protein correlates with L1 major capsid protein detection.
- The sole L1-containing transcript in HPV 11-infected tissue is a bicistronic E1-E4-L1 mRNA, with potential for dual protein expression.
Purpose of the Study:
- To establish whether the E1-E4 and L1 proteins can be expressed from the HPV 11 bicistronic E1-E4-L1 transcript.
- To investigate the expression levels and regulation of E1-E4 and L1 proteins from this transcript.
Main Methods:
- Cloning of the HPV 11 E1-E4-L1 sequence into expression vectors (p1393, pSPORT).
- Production of recombinant baculoviruses and infection of Sf9 cells.
- Analysis of protein expression using immunoblotting, ELISA, and in vitro transcription/translation assays.
- Site-directed mutagenesis and antisense oligonucleotide inhibition experiments.
Main Results:
- Recombinant baculovirus infection confirmed expression of both E1-E4 and L1 proteins.
- In vitro transcription/translation showed E1-E4 and L1 proteins produced at a 17:1 ratio from the bicistronic construct.
- Mutating the E1-E4 start codon or using antisense oligonucleotides modulated E1-E4 and L1 protein synthesis, indicating translational regulation.
Conclusions:
- The HPV 11 E1-E4-L1 bicistronic mRNA can indeed produce both E1-E4 and L1 proteins.
- Translational control mechanisms influence the expression ratio of these proteins, impacting viral assembly and infection.