Site-directed mutagenesis of Autographa californica nucleopolyhedrovirus (AcNPV) polyhedrin: effect on polyhedron structure.
Amino acids Lys34, His36, and Phe37 were substituted by PCR-mediated, site-directed mutagenesis for three Trp's in the AcNPV polyhedrin sequence. Phase contrast microscopy revealed refringent, amorphous polyhedra in the nuclei of infected cells. Electron microscopy confirmed a great variation in form and size of the mutated polyhedra. Although crystallization of the mutated polyhedrin occurred, it was irregular within each polyhedron. Virion occlusion was also severely affected. Virions were partially occluded, or only one virion was occluded per polyhedron. Results suggest that the substitution of these three amino acids affected the morphology of polyhedra, the uniformity of crystallization within each polyhedron, and the virion occlusion.
Bravo-Patiño
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Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del I.P.N., Irapuato, Gto. México.
Ibarra
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JE
D000595:Amino Acid Sequence; D001483:Base Sequence; D003460:Crystallization; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017924:Nucleopolyhedroviruses / Q000737:chemistry*; D000077183:Occlusion Body Matrix Proteins; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000737:chemistry*; D015678:Viral Structural Proteins
D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus