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Analysis of a temperature-sensitive mutation affecting the integration protein of Moloney murine leukemia virus
P L Schwartzberg1, M J Roth, N Tanese
1Department of Biochemistry, Columbia University College of Physicians and Surgeons, New York, New York 10032.
Abstract:
We have previously described a series of mutations in the 3' terminus of the pol gene of Moloney murine leukemia virus which adversely affect the establishment of an integrated provirus (Roth, M. J., Schwartzberg, P., Tanese, N., and Goff, S. P., J. Virol. 64, 4709-4717, 1990). While most of the mutations were unconditionally lethal for virus replication, we now demonstrate that one mutation, in6161-12, a 12-base pair linker insertion into the 3' end of the pol gene, causes a temperature-sensitive phenotype. This mutant virus is temperature-sensitive for IN protein function as well as viral replication.
Insights
A Moloney murine leukemia virus mutation causes temperature-sensitive replication and integrase (IN) protein function. This finding reveals a critical role for the pol gene
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Previous research identified mutations in the Moloney murine leukemia virus pol gene affecting provirus integration.
- Most prior mutations were lethal to viral replication.
Purpose of the Study:
- To investigate the impact of a specific pol gene mutation (in6161-12) on Moloney murine leukemia virus replication and integrase function.
- To characterize the temperature-sensitive phenotype associated with this mutation.
Main Methods:
- Site-directed mutagenesis to create a 12-base pair linker insertion (in6161-12) at the 3' end of the pol gene.
- Assessing viral replication and integrase (IN) protein function at different temperatures.
Main Results:
- The in6161-12 mutation confers a temperature-sensitive phenotype to the virus.
- Both viral replication and integrase (IN) protein function are impaired at the restrictive temperature.
Conclusions:
- The 3' end of the pol gene is crucial for Moloney murine leukemia virus replication and integrase function.
- The in6161-12 mutation provides a valuable tool for studying temperature-sensitive aspects of viral replication and IN protein activity.