NcRNA In UHRF1Complex Identification And Function Analysis

Across the whole human genome, about93%can be transcribed. However, most of them don’t correspond to annotated exons of proteins coding genes. These noncoding transcripts belong to divergent transcription. The promoter region of noncoding RNA is more conserved than the ones of the coding mRNA from evolutionary perspective. Furthermore, about more than half of ncRNAs are nuclear retention which means their potential roles in chromatin activity regulation in the nucleus.With the development of sequencing, amazing numbers of ncRNA including long ncRNA and small ncRNA are discovered. Both of them affect markedly the corresponding genes’expression. Moreover, researches about DNA methylation and hydroxymethylation, histone modification and chromatin remodeling that engage a series of chromatin-related complexes are flourishing. There are also studies showing that many chromatin related complexes such as PRC2and MLL1have direct interaction with ncRNA. In those chromatin associated ribonucleoprotein complex, ncRNA guides protein to targeted region, or ncRNA service as a scaffold to couple different complexes that cooperate to play a synergistic effect.What and which kinds of ncRNA would present in a complex? How do ncRNA and chromatin associated complexes regulate cooperatively chromatin activity and gene expression? What is spatio-temporal expression of ncRNA that anchor to complex in various development stages and different physiological states. Our research is aiming at these unsolved questions to search ncRNA that embed in the chromatin associated complex UHRF1, deduce their common features, establish interaction network and understand their mechanisms of action in regulating chromatin activity from a new angles. Our experiments show that UHRF1binds repeat-derived transcriptions which implies its function in regulate repeats’activity. UHRF1also participates in RNA processing at both protein and RNA level. Moreover, UHRF1involving in tRNA transcripts that with high frequency usage indicates its roles in tRNA gene expression and the evolution of tRNA gene copies. Interestingly, we found that UHRF1binds XIST and MALAT1, both of them are hotspot of ncRNA study which involve in X chromatin inactivation and tumors metastasis.Objective Searching for what and which kinds of ncRNA presenting in the chromatin associated complex UHRF1at a global level and studying further the function and underlying mechanism of these ncRNAs, which may provide a new insight into elucidate the function of UHRF1.Methods and results1、Applying TAP to purify UHRF1complex from UHRF1integrated HEK293T cell;2、Extracting RNA components from UHRF1complex and analyzing these ncRNA’s characteristic and potential function combining with protein and DNA components;3、Applying bioinformatics to establish the regulation network of ncRNA in UHRF1complex.ConclusionsWe find that UHRF1complex participate in RNA processing and influence activity of repeat-inclusive-chromatin. We also find that XIST and MALAT1are present in the UHRF1complex with high value of signal which indicates that UHRF1is associated with X chromatin inactivation and neoplasm metastasis.