Author: Wenbin Ji; Yibo Luo; Ejaz Ahmad; Song-Tao Liu
Title: Coordination between discrete Mitotic Arrest Deficient 1 (MAD1) domains is required for efficient mitotic checkpoint signaling Document date: 2017_11_1
ID: i4yquw4k_7
Snippet: When examined by Mis12 immunoprecipitation, in addition to MAD1 AA failing to recruit MAD2, slight reduction of steady levels of MAD2 binding to other MAD1 mutants were also noticed (Fig.1e) . The reduction of MAD2 binding became more obvious for some mutants when FLAG immunoprecipitation was performed using cells transfected with FLAG-MAD1 constructs (Fig 1f) . In addition, MAD1 ï„CTD and MAD1 ï„NTD cannot interact with endogenous MAD1, indica.....
Document: When examined by Mis12 immunoprecipitation, in addition to MAD1 AA failing to recruit MAD2, slight reduction of steady levels of MAD2 binding to other MAD1 mutants were also noticed (Fig.1e) . The reduction of MAD2 binding became more obvious for some mutants when FLAG immunoprecipitation was performed using cells transfected with FLAG-MAD1 constructs (Fig 1f) . In addition, MAD1 ï„CTD and MAD1 ï„NTD cannot interact with endogenous MAD1, indicating a dimerization defect (Fig 1e&f, note that mCherry-Mis12-MAD1 ï„NTD runs at the same position as endogenous MAD1. More on MAD1 dimerization in the last section of RESULTS). Taken together, results shown in Figure 1 confirm the important role of MAD1 MIM , but also reveal that both NTD and CTD of MAD1 are required for an efficient mitotic checkpoint.
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