ChIP-seq of HA-METTL14 in HepG2 cells

The METTL3-METTL14 heterodimer is the core component of the N6-methyltransferase complex (MTC) that catalyzes methylation of adenosine residues at the N(6) position of RNA. As the non-catalytic subunit of the MTC, METTL14 functions as the RNA-binding scaffold that recognizes the RNA substrate. We found METTL14 could interacte with chromatin through recognition of H3K36me3. ChIP-seq with HA-tagged METTL14 using an HA antibody identified 300 peaks in HepG2 cells, among which 53% peaks were located in the gene body and only 3% peaks were located in the promoter. The binding sites of METTL14 on chromatin correlate with that of H3K36me3.To investigat whether this interaction is Pol II dependent, we treated cells with a Pol II inhibitor 5,6-Dichlorobenzimidazole 1-β-D-ribofuranoside (DRB) for a short time. Interestingly, we found the interaction of METTL14 with chromatin in gene body region was not affected by DRB treatment . ChIP-seq using HA antibody in HepG2 cells with HA-METTL14 overexpression

METTL3-METTL14异二聚体是N6-甲基转移酶复合物（MTC）的核心组分，该复合物可催化RNA的N(6)位腺苷残基发生甲基化。作为MTC的非催化亚基，METTL14作为识别RNA底物的RNA结合支架发挥功能。我们发现，METTL14可通过识别H3K36me3与染色质发生相互作用。使用HA抗体对HA标记的METTL14进行染色质免疫沉淀测序（Chromatin Immunoprecipitation Sequencing, ChIP-seq），在HepG2细胞中共鉴定出300个结合峰，其中53%的峰位于基因体区域，仅3%的峰定位于启动子区域。METTL14在染色质上的结合位点与H3K36me3的结合位点具有相关性。为探究该相互作用是否依赖于RNA聚合酶II（Pol II），我们使用Pol II抑制剂5,6-二氯苯并咪唑-1-β-D-呋喃核糖苷（DRB）对细胞进行短时间处理。有趣的是，我们发现METTL14在基因体区域与染色质的相互作用并未受到DRB处理的影响。在过表达HA-METTL14的HepG2细胞中使用HA抗体开展染色质免疫沉淀测序实验。