Background: UBE2Q1-dependent ubiquitination of key proteins including β 1,4- galactosyltransferase (GalT1), and P53 might play a pivotal role in cancer development.
Objective: The present study aimed to evaluate the molecular analysis of possible interactions between UBE2Q1 with B4GALT1 and P53 proteins.
Methods: We established SW1116 colorectal cancer cell line stably transfected with UBE2Q1. To verify the overexpression of UBE2Q1, we performed western blot and fluorescent microscopy analysis. Using the immunoprecipitation (IP) product of the over-expressed protein on the silver staining gel, we observed the potential interacting partners of UBE2Q1. The Molecular Operating Environment (MOE) software was also used to perform the molecular docking of the UBC domain of UBE2Q1 (2QGX) with B4GALT1 (2AGD), and P53 (tetramerization (1AIE) and DNA binding domains (1GZH)) proteins.
Results: Western blot and IP analysis detected a UBE2Q1-GFP band in transfected cells, while no band was detected for mock-transfected cells. Moreover, the overexpression of UBE2Q1 tagged with GFP was observed under fluorescent microscopy as well with about 60-70% shining. Silver staining of IP gel revealed several bands in colorectal cancer (CRC) with UBE2Q1 overexpression. Protein- Protein interaction (PPI) analysis also depicted a high affinity of the UBC domain of UBE2Q1 to the B4GALT1 and P53 (tetramerization and DNA binding domains). Molecular docking also revealed hot-spot regions for all poses.
Conclusion: Our data suggest that UBE2Q1 as an E2 enzyme of ubiquitination system can interact with B4GALT1 and P53, and may contribute to the accumulation of misfolded important proteins and colorectal tumor development.
[http://dx.doi.org/10.1104/pp.105.067983] [PMID: 16339806]
[http://dx.doi.org/10.1007/s11033-020-05702-1] [PMID: 32770525]
[http://dx.doi.org/10.1007/s10735-014-9596-x] [PMID: 25311764]
[http://dx.doi.org/10.1002/mc.22747] [PMID: 29027712]
[http://dx.doi.org/10.2174/092986608783330396] [PMID: 18221015]
[http://dx.doi.org/10.2174/0929866527666200921164536] [PMID: 32957871]
[http://dx.doi.org/10.1007/s11033-021-06440-8] [PMID: 34086159]
[http://dx.doi.org/10.1002/gcc.21998] [PMID: 22927297]