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Control of membrane morphogenesis in bacteriophage PM2
Abstract:
The regulation of membrane formation in bacteriophage PM2 serves as a simple model for changes in membrane structure in eukaryotic cells. Prior to Pseudomonas host lysis, wild-type virions mature to an icosahedral morphology at the inner face of the cytoplasmic membrane. The preliminary characterization of two temperature-sensitive mutants of PM2 is described. In cells infected at the restrictive temperature with ts 1, an abundance of "empty" virus-size membrane vesicles are seen. Synthesis of DNA is also reduced in ts 1 infected cells. The preponderance of vesicles is not seen in cells infected with wild-type virus or with ts 1 at the permissive temperature. The "empty" appearance of the viral membranes suggests that viral DNA is not encapsulated. The major viral capsid protein (MW 26,000) is located just outside the viral membrane and normally sidiments with host and virus membranes. This protein made by mutant ts 5 does not pellet with these membranes; instead, large amounts of capsid protein can be precipitated from the supernatant with TCA. Compared to cells infected with wild type virus, cells infected with ts 5 at the restrictive temperature produced inside the cell an abundance of virus-size membrane vesicles. Taken together, these results with viral mutants suggest that formation of a viral membrane of the proper size does not require a DNA core around which to form, or an outer scaffolding of coat protein against which to form a spherical bilayer.
Insights
Bacteriophage PM2 mutants reveal that viral membrane formation does not require DNA or coat protein scaffolding. This finding offers insights into membrane biogenesis in both viruses and eukaryotic cells.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Bacteriophage PM2 membrane formation provides a model for eukaryotic membrane dynamics.
- Wild-type virions mature at the inner face of the Pseudomonas host cytoplasmic membrane before lysis.
Purpose of the Study:
- To characterize two temperature-sensitive mutants of bacteriophage PM2.
- To investigate the roles of DNA and capsid protein in viral membrane formation.
Main Methods:
- Analysis of temperature-sensitive mutants (ts 1 and ts 5) of bacteriophage PM2.
- Observation of viral morphology and membrane vesicle formation in infected Pseudomonas cells at restrictive and permissive temperatures.
- Biochemical analysis of viral capsid protein pelleting with membranes.
Main Results:
- Mutant ts 1 produced abundant, empty virus-size membrane vesicles and reduced DNA synthesis at restrictive temperatures.
- Mutant ts 5 showed defective capsid protein association with membranes and produced virus-size membrane vesicles.
- The formation of correctly sized viral membranes was observed even without DNA encapsulation or complete coat protein scaffolding.
Conclusions:
- Viral membrane formation can occur independently of a DNA core.
- The assembly of viral membranes does not necessitate outer coat protein scaffolding.
- These findings contribute to understanding fundamental principles of membrane biogenesis.