JMJD6 and U2AF65 co-regulate alternative splicing in both JMJD6 enzymatic activity dependent and independent manner

Abstract

JMJD6, a jumonji C (Jmj C) domain-containing protein demethylase and hydroxylase, has been implicated in an array of biological processes. It has been shown that JMJD6 interacts with and hydroxylates multiple serine/arginine-rich (SR) proteins and SR related proteins, including U2AF65, all of which are known to function in alternative splicing regulation. However, whether JMJD6 is widely involved in alternative splicing and the molecular mechanism underlying JMJD6-regulated alternative splicing have remained incompletely understood. Here, by using RASL-Seq, we investigated the functional impact of RNA-dependent interaction between JMJD6 and U2AF65, revealing that JMJD6 and U2AF65 co-regulated a large number of alternative splicing events. We further demonstrated the JMJD6 function in alternative splicing in jmjd6 knockout mice. Mechanistically, we showed that the enzymatic activity of JMJD6 was required for a subset of JMJD6-regulated splicing, and JMJD6-mediated lysine hydroxylation of U2AF65 could account for, at least partially, their co-regulated alternative splicing events, suggesting both JMJD6 enzymatic activity-dependent and independent control of alternative splicing. These findings reveal an intimate link between JMJD6 and U2AF65 in alternative splicing regulation, which has important implications in development and disease processes.

JMJD6 Licenses ERα-Dependent Enhancer and Coding Gene Activation by Modulating the Recruitment of the CARM1/MED12 Co-activator Complex

“…S1H and S1I). It should be noted that JMJD6 oligomerization remained intact even under denature conditions, which was consistent with previous reports (Liu et al, 2013, Yi et al, 2017, Han et al, 2012). Unlike JMJD6 binding sites being induced robustly, those 133 JMJD6 binding sites lost in response to estrogen treatment were either with low tag density at both conditions, with or without E 2 treatment, but however were falsely predicted as peaks, or the reduction of tag density were very subtle.…”

“…S5G). It should be noted that JMJD6 oligomerization was lost when histidine 187 was mutated to alanine, which was consistent with previous reports (Yi et al, 2017, Liu et al, 2013, Han et al, 2012). Taken together, our data suggested that JMJD6 is required for estrogen-induced MCF7 breast cancer cell and tumor growth, which is dependent on its enzymatic activity.…”

“…We recently reported a transcriptional paradigm in which JMJD6 regulates promoter-proximal Pol II pausing release in a distal manner (Liu et al, 2013). Besides its function in transcriptional control, we and others have shown that JMJD6 also interacts with multiple splicing factors, and is involved in gene splicing control (Heim et al, 2014, Webby et al, Liu et al, 2013, Rahman et al, 2011, Yi et al, 2017). JMJD6 has been implicated in a multitude of biological processes, including embryonic development (Bose et al, 2004, Li et al, 2003, Kunisaki et al, 2004), cell cycle control (Wang et al, 2014a), cellular proliferation and motility (Lee et al, 2012, Chen et al, 2014), adipocyte differentiation (Hu et al, 2015) and development of various types of cancers, such as breast (Poulard et al, 2015, Lee et al, 2012, Aprelikova et al, 2016), lung (Zhang et al, 2013) and colon cancer (Wang et al, 2014a).…”

Highly multiplexed oligonucleotide probe-ligation testing enables efficient extraction-free SARS-CoV-2 detection and viral genotyping

“…While we have not explicitly tested higher levels of ligation probe multiplexing in this study, previous RASL-seq studies have employed panels of >5,000 probe sets. 27 With appropriate design of non-interfering capture and ligation probe sets, we thus anticipate that we could achieve multiplexing of up to 10,000 probe sets, without technical artifacts.…”

Jumonji domain‐containing protein 6 protein and its role in cancer

“…JMJD6 interacts with U2AF65 that binds to Flt1 mRNA . More recently, it was reported that JMJD6 and U2AF65 directly bind to pre‐mRNA and coregulate a large set of alternative splicing events …”