Interaction Analysis Of PRC2 Components And Interaction Screening Of Its Antagonistic Proteins JMJs

In plants and animals,Polycomb Group(PcG)protein plays an important role in regulating gene expression and maintaining developmental status.PcG proteins assemble into various complexes,and the main formsarePolycomb repressive complex 1(PRCl)and Polycomb repressive complex 2(PRC2).PRC2,a four-subunit histone methyltransferase complex,acts to inhibit gene expression.In the process of individual growth and development,affected by both environment and gene,gene expression regulation is not completed by a single protein,and its function and interaction mechanism are more complex with the increase of PRC2 components copy numbers.The PRC2 components copy number of the tree model plant Populus trichocarpa is more than that of the herbaceous model plant Arabidopsis thaliana,and the interaction relationship is not clear.So,in this study,PRC2 components of Arabidopsis thaliana and Populus trichocarpa were cloned to construct into yeast two-hybrid vector,and the interaction between PRC2 components of model plant Arabidopsis thaliana and PRC2 components of Populus trichocarpa was verified by yeast two-hybrid assay.The Jumonji(JMJ)proteins,antagonizing PRC2 action,have H3K27 demethylase activity in Arabidopsis thaliana.These genes were shown to be involved in epigenetic reprogramming,flowering time control,brassinosteroid signalling and circadian clock.However,relevant interactions have not been reported at present,so we screened the JMJ 13 interacting proteins through yeast two-hybrid library screening assay,laying a foundation for exploring its function in plant development.Results are as follows:(1)there were new interactions of PRC2 components of Arabidopsis thaliana.(2)there were differences in the interaction between the PRC2 components of Populus trichocarpa and that of Arabidopsis thaliana.(3)57 interacting proteins were screened out from 4191 clones by yeast two-hybrid screening library,which were involved in root development,stress resistance response,auxin regulation,protein ubiquitination modification,stamen development,pollen germination.(4)Transcription factors MYB3,MYB4,MYB32,and GTE4 were found,and the interaction with JMJ 13 was further verified by yeast two-hybrid.