The Study Of Regulation Of INO80 Chromatin Remodeling Complex On The Expression Of ANKRD1

INO80 complex is an important member of SWI/SNF family;it is a significant chromatin remodeling complex in the cell,it can regulate many genes' expression.The complex has a very complicated structure,and was formed by several subunits,including ARP8 YY1 and so on.INO80 is a chromatin remodeling complex depending on the ATPase.When there is ATP provides energy in the cell,the complex can make nucleosome to slide along the DNA strand.So that it can change the accessibility of DNA strand.It can also replace the variant histone H2 AZ out of histone octamer to change the structure of chromatin.By these two ways,INO80 chromatin remodeling complex can transform the chromatin structure to regulate genes' transcription.There are three kinds of proteins in the MARP family,including ANKRD1 ANKRD2 and ANKRD23;they are all highly conserved in mammals.ANKRD1 is a kind of embryonic gene.It can be highly expressed in the growth of embryo and fetus,and it will gradually reduce as we grow older.ANKRD1 have many biological effects in regulating the development of the heart.Also,many clinical experiments proved that ANKRD1 is highly expressed in cardiac hypertrophy and heart failure patient.It is a new land marker of these diseases.In the previous experiment,we knocked down INO80 in Hela cells.Then we sent the sample to do RNA-seq.From the results,we can find expression vibration of many genes,which including ANKRD1.So that,we guessed that the expression of ANKRD1 can be regulated by INO80.To prove this,we knocked down INO80 and its subunit ARP8 to observe the expression of ANKRD1.The results showed that the expression of ANKRD1 elevated both in protein level and mRNA level.Furthermore we tried to find the appropriate conditions to do ChIP experiment in Hela cells.With the conditions,we used INO80 antibody to conduct the ChIP experiment.The result shows there was a occupy peak of INO80 in the promoter region of ANKRD1.In the following research,we constructed the plasmid pcDNA3.1-Flag-ANKRD1 incase for other experiments.