Structural role of the matrix protein of type D retroviruses in gag polyprotein stability and capsid assembly.
To obtain a better understanding of the role of the gag gene-encoded matrix (MA) protein in the assembly and maturation of type D retroviruses, we have made five mutants with specific in-frame deletions within the p10-coding region by the use of oligonucleotide-directed mutagenesis. The changes in the Gag polyprotein made by these mutations resulted in almost identical phenotypes. In cells expressing mutant genomes, the mutant Gag polyproteins were synthesized and modified with myristic acid in a normal manner. However, they were so unstable that the bulk of the newly synthesized polyproteins was degraded within 1 h without being processed into mature structural polypeptides. In contrast, wild-type polyproteins have a processing half-life of 3.0 to 3.5 h. The mutant Gag polyproteins were assembled with very low efficiency into capsids in the cytoplasm of the mutant-infected cells. Moreover, the few capsids that formed were neither released from nor accumulated in the cells. These results suggest that the matrix protein plays an important role in guiding the correct folding of the Gag polyprotein, which is presumably crucial for both stabilizing the molecule and facilitating the intermolecular interactions that occur during assembly of immature capsids.
Rhee
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SS
Department of Microbiology, University of Alabama, Birmingham 35294.
Hunter
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E
D000595:Amino Acid Sequence; D002213:Capsid / Q000235:genetics*; D002872:Chromosome Deletion; D015683:Gene Products, gag / Q000235:genetics*; D005814:Genes, Viral; D015750:Genes, gag; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism; D015345:Oligonucleotide Probes; D010641:Phenotype; D012190:Retroviridae / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification
D015683:Gene Products, gag; D009227:Myristic Acids; D015345:Oligonucleotide Probes; D014764:Viral Proteins; D019814:Myristic Acid