Screen And Identify Protein Interacts With MCM4C-terminal Domain (MCM4-CTD) By Yeast Two-hybrid System

The stringency and accuracy of DNA replication is very important for thetransmission of genetic information. A hexamer minichromosome maintenance proteinscomplex, which is composed of six subunits, plays a role as a helicase in replication ofeukaryote cells. When DNA damage occurs, replication forks become stalled, and theexcessive unwinding produces overmuch single stranded DNA (ssDNA), which leads theinstability of replication forks even all the genome. The research reveals that MCM4C-terminal domain of MCM complex can prevent excessive unwinding in case ofovermuch production of single stranded DNA at stalled replication forks, which indicatesthat MCM4-CTD has a important role in DNA damage response process when DNA isdamage, and the coding gene of MCM4-CTD is648bp. With the yeast two-hybrid system,we clone MCM4-CTD into BD vector, and screen and identify a protein interact with itfrom yeast cDNA library, SNF12, which is a subunit of Chromatin remodeling complexSWI/SNF. So we presume that chromatin remodeling activity participates in regulation ofDNA replication in the DNA damage response progress.