CARM1 Methylates Chromatin Remodeling Factor BAF155 to Enhance Tumor Progression and Metastasis

Wang, Lu; Zhao, Zibo; Meyer, Mark B.; Saha, Sandeep; Yu, Menggang; Guo, Ailan; Wisinski, Kari B.; Huang, Wei; Cai, Weibo; Pike, J. Wesley; Yuan, Ming; Ahlquist, Paul; Xu, Wei

doi:10.1016/j.ccr.2013.12.007

Cited by 219 publications

(260 citation statements)

References 37 publications

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“…Several of these N-terminal residues are predicted to be sites of posttranslational modifications, including multiple arginines that represent potential targets of the arginine methyltransferase enzymes PRMT1, PRMT4, and PRMT5 (31,32). We, and others, have shown that myeloid differentiation is regulated by specific arginine methylation events, and PRMT4 modifies several proteins in the larger DPF2 interactome, including RUNX1 and the SWI/SNF subunit BAF155 (8,33). Thus, the recognition of histone modifications by the PHD domains of DPF2 potentially integrates multiple epigenetic inputs and likely enables the targeting of DPF2 to specific chromatin sites.…”

Section: Discussionmentioning

confidence: 99%

Histone-binding of DPF2 mediates its repressive role in myeloid differentiation

Huber

Greenblatt

Davenport

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Double plant homeodomain finger 2 (DPF2) is a highly evolutionarily conserved member of the d4 protein family that is ubiquitously expressed in human tissues and was recently shown to inhibit the myeloid differentiation of hematopoietic stem/progenitor and acute myelogenous leukemia cells. Here, we present the crystal structure of the tandem plant homeodomain finger domain of human DPF2 at 1.6-Å resolution. We show that DPF2 interacts with the acetylated tails of both histones 3 and 4 via bipartite binding pockets on the DPF2 surface. Blocking these interactions through targeted mutagenesis of DPF2 abolishes its recruitment to target chromatin regions as well as its ability to prevent myeloid differentiation in vivo. Our findings suggest that the histone binding of DPF2 plays an important regulatory role in the transcriptional program that drives myeloid differentiation.T he founding member of the d4 family of proteins, d4, zinc, and double plant homeodomain (PHD) finger 2 (DPF2, also known as requiem/REQ or ubi-d4), was initially discovered as a factor required for apoptosis in myeloid cells (1). d4 proteins, which in humans also include DPF1 and DPF3b, are characterized by an N-terminal requiem domain, a central C2H2-type zinc finger domain, and a C-terminal tandem PHD finger (2). PHD fingers, which contain two zinc finger motifs, are notable for their ability to read a diverse number of posttranslational modifications, including unmodified, methylated, or acetylated lysines, as well as unmodified arginines (3). Besides such putative binding capabilities of the DPF2 tandem PHD finger domain, relatively little is known about the regulation and function of DPF2 or its remaining individual domains. Previous studies have shown that DPF2 bridges SWI/SNF components and RelB/p52 to affect noncanonical NF-κB signaling (4), acts as a globin switching factor (5), and is a target for Staufen-1-mediated mRNA decay (5). Notably, DPF2 is expressed ubiquitously in human tissues compared with DPF1 and DPF3b (6, 7). DPF2, along with DPF1 and DPF3b, has been implicated in a range of human cancers, including cervical cancer and acute myelogenous leukemia (AML) (8-13). Runt-related transcription factor 1 (RUNX1, also known as AML1) functions as an AML tumorsuppressor gene, which is frequently inactivated through somatic mutations and chromosomal translocations, including t(8;21), which produces the AML1-ETO fusion protein (14). Recent work has shown that recruitment of DPF2 into a RUNX1-containing repressor complex inhibits the expression of RUNX1 target genes, including the myeloid-specific microRNA miR-223, and inhibits myeloid differentiation (8). DPF2 recruitment appears to depend on arginine methylation events of RUNX1, as it is blocked by mutation of RUNX1 Arg223 or chemical inhibition of the type I arginine methyltransferase PRMT4. Knockdown of either DPF2 or PRMT4 increases miR-223 gene expression and myeloid differentiation. These findings suggest a model in which DPF2 and RUNX1 form a methylation-dependent repressive co...

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Section: Discussionmentioning

confidence: 99%

Histone-binding of DPF2 mediates its repressive role in myeloid differentiation

Huber

Greenblatt

Davenport

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…In the present study, Halo-tag and Flag-tag CARM1 (WT and QM) were expressed in HEK293T cells with CARM1 knocked out by ZFN (HEK293T CARM1 −/− ) [28]. Immunoprecipitation was performed using Anti-FLAG ® M2 Beads.…”

Section: O-glcnacylation Does Not Affect Cellular Localization Stabimentioning

confidence: 99%

“…CARM1-null human embryonic kidney (HEK293T), MCF7 (ERa positive breast cancer cell line) and MDA-MB-231 (triplenegative breast cancer cell line) cell lines were described in [28]. CARM1-null Cal51 cell lines were generated using the same method as described in [28].…”

Section: Generation Of Carm1 (Wt or Qm) Stably Expressing Cell Linesmentioning

confidence: 99%

“…We have previously reported CARM1 knockout MDA-MB-231 (MDA − MB − 231 CARM1 −/− ) and MCF7 (MCF7 CARM1 −/− ) cell lines generated by zinc finger nuclease (ZFN) technology [28]. Flag-tagged CARM1…”

Section: O-glcnacylation Does Not Affect Cellular Localization Stabimentioning

confidence: 99%

“…CARM1-null Cal51 cell lines were generated using the same method as described in [28]. Flag-tagged mouse CARM1…”

Section: Generation Of Carm1 (Wt or Qm) Stably Expressing Cell Linesmentioning

confidence: 99%

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O-GlcNAcylation of co-activator-associated arginine methyltransferase 1 regulates its protein substrate specificity

Charoensuksai

Kühn

Wang

et al. 2015

Biochemical Journal

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Co-activator-associated arginine methyltransferase 1 (CARM1) asymmetrically di-methylates proteins on arginine residues. CARM1 was previously known to be modified through O-linked-β-N-acetylglucosaminidation (O-GlcNAcylation). However, the site(s) of O-GlcNAcylation were not mapped and the effects of O-GlcNAcylation on biological functions of CARM1 were undetermined. In the present study, we describe the comprehensive mapping of CARM1 post-translational modification (PTM) using top-down MS. We found that all detectable recombinant CARM1 expressed in human embryonic kidney (HEK293T) cells is automethylated as we previously reported and that about 50% of this automethylated CARM1 contains a single O-linked-β-N-acetylglucosamine (O-GlcNAc) moiety [31]. The O-GlcNAc moiety was mapped by MS to four possible sites (Ser595, Ser598, Thr601 and Thr603) in the C-terminus of CARM1. Mutation of all four sites [CARM1 quadruple mutant (CARM1QM)] markedly decreased O-GlcNAcylation, but did not affect protein stability, dimerization or cellular localization of CARM1. Moreover, CARM1QM elicits similar co-activator activity as CARM1 wild-type (CARM1WT) on a few transcription factors known to be activated by CARM1. However, O-GlcNAc-depleted CARM1 generated by wheat germ agglutinin (WGA) enrichment, O-GlcNAcase (OGA) treatment and mutation of putative O-GlcNAcylation sites displays different substrate specificity from that of CARM1WT. Our findings suggest that O-GlcNAcylation of CARM1 at its C-terminus is an important determinant for CARM1 substrate specificity.

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High affinity binding of H3K14ac through collaboration of bromodomains 2, 4 and 5 is critical for the molecular and tumor suppressor functions of PBRM1

et al. 2019

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Polybromo‐1 ( PBRM 1) is an important tumor suppressor in kidney cancer. It contains six tandem bromodomains ( BD s), which are specialized structures that recognize acetyl‐lysine residues. While BD 2 has been found to bind acetylated histone H3 lysine 14 (H3K14ac), it is not known whether other BD s collaborate with BD 2 to generate strong binding to H3K14ac, and the importance of H3K14ac recognition for the molecular and tumor suppressor function of PBRM 1 is also unknown. We discovered that full‐length PBRM 1, but not its individual BD s, strongly binds H3K14ac. BD s 2, 4, and 5 were found to collaborate to facilitate strong binding to H3K14ac. Quantitative measurement of the interactions between purified BD proteins and H3K14ac or nonacetylated peptides confirmed the tight and specific association of the former. Interestingly, while the structural integrity of BD 4 was found to be required for H3K14ac recognition, the conserved acetyl‐lysine binding site of BD 4 was not. Furthermore, simultaneous point mutations in BD s 2, 4, and 5 prevented recognition of H3K14ac, altered promoter binding and gene expression, and caused PBRM 1 to relocalize to the cytoplasm. In contrast, tumor‐derived point mutations in BD 2 alone lowered PBRM 1's affinity to H3K14ac and also disrupted promoter binding and gene expression without altering cellular localization. Finally, overexpression of PBRM 1 variants containing point mutations in BD s 2, 4, and 5 or BD 2 alone failed to suppress tumor growth in a xenograft model. Taken together, our study demonstrates that BD s 2, 4, and 5 of PBRM 1 collaborate to generate high affinity to H3K14ac and tether PBRM 1 to chromatin. Mutations in BD 2 alone weaken these interactions, and this is sufficient to abolish its molecular and tumor suppressor functions.

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CARM1 Methylates Chromatin Remodeling Factor BAF155 to Enhance Tumor Progression and Metastasis

Cited by 219 publications

References 37 publications

Histone-binding of DPF2 mediates its repressive role in myeloid differentiation

Histone-binding of DPF2 mediates its repressive role in myeloid differentiation

O-GlcNAcylation of co-activator-associated arginine methyltransferase 1 regulates its protein substrate specificity

High affinity binding of H3K14ac through collaboration of bromodomains 2, 4 and 5 is critical for the molecular and tumor suppressor functions of PBRM1

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