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The E2 "gene" of bovine papillomavirus encodes an enhancer-binding protein | JoVE Visualize

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The E2 "gene" of bovine papillomavirus encodes an enhancer-binding protein

C Moskaluk, D Bastia

Proceedings of the National Academy of Sciences of the United States of America|March 1, 1987

View abstract on PubMed

Summary

Researchers created a hybrid protein from bovine papillomavirus-1 E2 gene and beta-galactosidase. This protein binds to specific DNA sequences in the bovine papillomavirus enhancer region, revealing a consensus binding motif.

Area of Science:

Molecular biology
Virology
Biochemistry

Background:

Bovine papillomavirus-1 (BPV-1) is a virus that can cause tumors.
The E2 early open reading frame (gene) of BPV-1 is crucial for viral replication and transcription.
Understanding the DNA-binding properties of viral proteins is essential for developing antiviral strategies.

Purpose of the Study:

To investigate the DNA-binding specificity of the BPV-1 E2 protein.
To identify the consensus DNA sequence recognized by the BPV-1 E2 protein within the viral enhancer region.

Main Methods:

Fusion of the BPV-1 E2 gene with the beta-galactosidase gene in the pJG200 vector.
Expression and partial purification of the hybrid protein from Escherichia coli.
DNase I-cleavage protection analysis to map DNA-protein interactions.

Main Results:

The hybrid E2-beta-galactosidase protein specifically bound to the enhancer region of BPV-1 DNA.
DNase I protection analysis identified a specific DNA sequence bound by the protein.
A consensus binding motif (AATTGGCGGNNCG) and a palindromic sequence (ATCGGTG/CACCGAT) were determined.

Conclusions:

The BPV-1 E2 protein recognizes a specific DNA consensus sequence within the viral enhancer.
This binding is critical for regulating viral gene expression and replication.
The identified consensus sequence provides insights into the molecular mechanisms of BPV-1 pathogenesis.

Biomedical Engineering

Biomechanical Engineering

The E2 "gene" Of Bovine Papillomavirus Encodes An Enhancer-binding Protein