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Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
Published on: March 22, 2016
Moloney murine leukemia virus integrase and reverse transcriptase interact with PML proteins.
Yuuki Okino1, Yujin Inayoshi, Yusuke Kojima
1Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya, 464-8603, Japan.
Journal of Biochemistry
|June 12, 2012
Summary
Moloney murine leukemia virus integrase and reverse transcriptase interact with PML proteins. PML proteins are essential for efficient viral cDNA synthesis and integration, suggesting they activate reverse transcription.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Moloney murine leukemia virus (MMLV) is a retrovirus.
- PML proteins are known to be involved in various cellular processes.
- The role of PML proteins in retroviral replication is not fully understood.
Purpose of the Study:
- To investigate the interaction between MMLV integrase and reverse transcriptase with PML proteins.
- To determine the functional significance of this interaction in viral replication.
Main Methods:
- Pull-down assays and co-immunoprecipitation were used to study protein interactions.
- Immunofluorescence microscopy was employed to assess co-localization.
- Small interfering RNA (siRNA) was used to knock down PML expression.
Main Results:
- Both MMLV integrase and reverse transcriptase were found to interact with PML proteins in cell extracts and infected cells.
- Integrase co-localized with PML proteins, potentially within PML bodies.
- SUMOylation of integrase by PIASy and SUMO proteins did not affect its interaction with PML.
- Knockdown of PML resulted in decreased viral cDNA levels and integration efficiency.
Conclusions:
- PML proteins interact with key MMLV enzymes, integrase and reverse transcriptase.
- PML proteins play a crucial role in promoting MMLV reverse transcription and integration.
- PML proteins appear to activate reverse transcription during MMLV infection.
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