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Published on: April 21, 2015
Identification of proteins that interact with murine cytomegalovirus early protein M112-113 in brain
Hui Wang1, Xing-Lou Liu, Sai-Nan Shu
1Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Background:
Murine cytomegalovirus (MCMV) early protein M112-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis. To investigate the biological function of M112-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus (CMV), a screening for proteins interacting with M112-113 was performed by a yeast two-hybrid system.
Methods:
Bait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast. After confirmation of the expression of MCMV M112-113 in yeast, the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M112-113. Interactions between M112-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion.
Results:
Two proteins interacting with M112-113 were identified, including metastasis-associated 1 (MTA1) and zinc finger, CCHC domain containing 18 (ZCCHC18). M112-113 protein could interact with MTA1 or ZCCHC18 in yeast and mammalian cells.
Conclusion:
The interactions of M112-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system.
Insights
Murine cytomegalovirus (MCMV) protein M112-113 interacts with metastasis-associated 1 (MTA1) and ZCCHC18. These interactions may contribute to MCMV-associated central nervous system disease pathogenesis.
Area of Science:
- Virology
- Neuroscience
- Molecular Biology
Background:
- Murine cytomegalovirus (MCMV) early protein M112-113 is crucial for viral DNA replication and pathogenesis.
- Cytomegalovirus (CMV) can cause brain disorders, but the role of M112-113 in this process is unclear.
Purpose of the Study:
- To investigate the biological function of MCMV M112-113 protein in the pathogenesis of CMV-induced brain disorders.
- To identify proteins that interact with M112-113 using a yeast two-hybrid system.
Main Methods:
- Constructed bait plasmid pGBKT7-M112-113 and transformed into AH109 yeast.
- Screened a mouse brain cDNA library for interacting proteins using yeast two-hybrid assay.
- Verified interactions via yeast two-hybrid and chemiluminescent co-immunoprecipitation in yeast and mammalian cells.
Main Results:
- Identified two interacting proteins: metastasis-associated 1 (MTA1) and zinc finger, CCHC domain containing 18 (ZCCHC18).
- Confirmed M112-113 interacts with MTA1 and ZCCHC18 in both yeast and mammalian cells.
Conclusions:
- The interactions between MCMV M112-113 and MTA1 or ZCCHC18 are potentially linked to the pathogenesis of MCMV-associated central nervous system diseases.
