Expression profiling by high throughput sequencing Genome binding/occupancy profiling by high throughput sequencing Other
Summary
N6-methyladenosine (m6A) plays critical roles in a wide range of physiological and pathological processes, and m6A methyltransferases are still being discovered for different RNA species in mammals. Here report a new m6A methyltransferase ZCCHC4 that methylates ribosomal RNA (rRNA) and interacts with messenger RNA (mRNA), ZCCHC4 has methylation activity on rRNA with a preference for AAC consensus sequence. ZCCHC4 can affect global translation through m6A methylation, mostly on 28S rRNA.
Overall design
We used PAR-CLIP to identify the binding targets of ZCCHC4 in HeLa cells. We used polysome sequencing to identify targets of ZCCHC4 wild type and knock out HepG2 cell for translation. We used m6A-seq to identify changes in m6A and RNA expression ZCCHC4 KO HepG2 cells.