The yeast two-hybrid screen for Ndt80-interacting proteins
Date Issued
2006
Date
2006
Author(s)
Ho, Chia-Wei
DOI
en-US
Abstract
In budding yeast Saccharomyces cerevisiae, cells defective in meiotic recombination and chromosome synapsis undergo checkpoint-mediated arrest at the pachytene stage. Previous study suggests that this checkpoint-mediated arrest is through Ndt80. Ndt80 is a meiosis-specific transcription factor that activates the expression of middle and late sporulation genes, including genes required for nuclear division and spore formation. Ndt80 is regulated by pachytene checkpoint at transcriptional and post-translational level. However, the detail mechanism is not clear. In our previous study, we found a dominant allele of Ndt80, Ndt80-bc. The deletion region of Ndt80-bc is in 346~402 a.a. We hypothesized that this region is the binding target of pachytene checkpoint to Ndt80. We proposed that the pachytene checkpoint regulates the activity of Ndt80 through protein-protein interaction.
In this study, we perform yeast two-hybrid screen to identify potential Ndt80-interacting proteins. We find that Akr2 has the potential to interact with Ndt80. Moreover, overproduction of Ndt80 and Akr2 in zip1 mutant decreases the suppression effect of Ndt80 overproduction in zip1 mutant. We suppose that Akr2 may negatively regulate Ndt80 activity. However, zip1akr2 mutant is still arrest at pachytene stage, suggesting that Akr2 may not be the most critical factor that negatively regulates Ndt80 activity. Furthermore, Akr2 also interacts with Ndt80-bc, suggesting that the different activity of Ndt80 and Ndt80-bc is not caused by Akr2, and there should be other reason that leads to the difference. We suggest that there would be other factors involved in the control mechanism.
Subjects
粗絲期檢控點
孢子
酵母菌雙雜合系統
減數分裂
pachytene checkpoint
yeast two-hybrid screen
meiosis
Ndt80
sporulation
Type
other