CHD3 and CHD4 form distinct NuRD complexes with different yet overlapping functionality

Hoffmeister, Helen; Fuchs, Andreas; Erdel, Fabian; Pinz, Sophia; Gröbner-Ferreira, Regina; Bruckmann, Astrid; Deutzmann, Rainer; Schwartz, Uwe; Maldonado, R.; Huber, Claudia; Dendorfer, Anne-Sarah; Rippe, Karsten; Längst, Gernot

doi:10.1093/nar/gkx711

Cited by 76 publications

(97 citation statements)

References 119 publications

(154 reference statements)

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“…Corroborating these data, recent findings by Hoffmeister and colleagues provide strong biochemical evidence demonstrating that mammalian CHD3 and CHD4 also form isoform-specific NuRD complexes (2017). Perhaps most compelling is the discovery that these distinct CHD3 and CHD4-NuRD containing complexes are also independently recruited to sites of induced DNA damage, suggesting overlapping functionality in this context (Hoffmeister et al, 2017). These data provide a particularly fitting explanation for the phenotypes we observed in…”

Section: (Ok615);let-418(n3536) Mutants Versus Rad-54(ok615) Alonementioning

confidence: 59%

Maintenance of Genome Integrity by Mi2 Homologs CHD-3 and LET-418 in Caenorhabditis elegans

et al. 2018

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Meiotic recombination depends upon the tightly coordinated regulation of chromosome dynamics and is essential for the production of haploid gametes. Central to this process is the formation and repair of meiotic double-stranded breaks (DSBs), which must take place within the constraints of a specialized chromatin architecture. Here, we demonstrate a role for the nucleosome remodeling and deacetylase (NuRD) complex in orchestrating meiotic chromosome dynamics in Caenorhabditis elegans. Our data reveal that the conserved Mi2 homologs Chromodomain helicase DNA binding protein (CHD-3) and its paralog LET-418 facilitate meiotic progression by ensuring faithful repair of DSBs through homologous recombination. We discovered that loss of either CHD-3 or LET-418 results in elevated p53-dependent germline apoptosis, which relies on the activation of the conserved checkpoint kinase CHK-1. Consistent with these findings, chd-3 and let-418 mutants produce a reduced number of offspring, indicating a role for Mi2 in forming viable gametes. When Mi2 function is compromised, persisting recombination intermediates are detected in late pachytene nuclei, indicating a failure in the timely repair of DSBs. Intriguingly, our data indicate that in Mi2 mutant germ lines, a subset of DSBs are repaired by non-homologous end joining, which manifests as chromosomal fusions.We find that meiotic defects are exacerbated in Mi2 mutants lacking CKU-80, as evidenced by increased recombination intermediates, corpses, and defects in chromosomal integrity. Taken together, our findings support a model wherein the C. elegans Mi2 complex maintains genomic integrity through reinforcement of a chromatin landscape suitable for homology-driven repair mechanisms.4

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Section: (Ok615);let-418(n3536) Mutants Versus Rad-54(ok615) Alonementioning

confidence: 59%

Maintenance of Genome Integrity by Mi2 Homologs CHD-3 and LET-418 in Caenorhabditis elegans

et al. 2018

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“…The pPCRScript_slog l-gla75, encoding the nucleosome positioning sequence, used for the assembly reaction (see below), was described in detail in Ref. 49. The pT11 plasmid DNA, used for stopping remodeling reactions (see below), was obtained from Dr. Joachim Griesenbeck (Biochemistry III, University of Regensburg, Regensburg, Germany).…”

Section: Plasmids and Constructsmentioning

confidence: 99%

“…The purification for C-terminal FLAG-tagged human wtBRG1 and its mutant proteins was adapted from the protocol in Ref. 49. In brief, human wtBRG1 and its Q and I motif mutants were purified from 1-2 ϫ 10 8 SF21 cells.…”

Section: Purification Of Flag-tagged Brg1 Proteins From Insect Cellsmentioning

confidence: 99%

Elucidation of the functional roles of the Q and I motifs in the human chromatin-remodeling enzyme BRG1

Hoffmeister

Fuchs

Strobl

et al. 2019

Journal of Biological Chemistry

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The Snf2 proteins, comprising 53 different enzymes in humans, belong to the SF2 family. Many Snf2 enzymes possess chromatin-remodeling activity, requiring a functional ATPase domain consisting of conserved motifs named Q and I-VII. These motifs form two recA-like domains, creating an ATPbinding pocket. Little is known about the function of the conserved motifs in chromatin-remodeling enzymes. Here, we characterized the function of the Q and I (Walker I) motifs in hBRG1 (SMARCA4). The motifs are in close proximity to the bound ATP, suggesting a role in nucleotide binding and/or hydrolysis. Unexpectedly, when substituting the conserved residues Gln 758 (Q motif) or Lys 785 (I motif) of both motifs, all variants still bound ATP and exhibited basal ATPase activity similar to that of wildtype BRG1 (wtBRG1). However, all mutants lost the nucleosome-dependent stimulation of the ATPase domain. Their chromatin-remodeling rates were impaired accordingly, but nucleosome binding was retained and still comparable with that of wtBRG1. Interestingly, a cancer-relevant substitution, L754F (Q motif), displayed defects similar to the Gln 758 variant(s), arguing for a comparable loss of function. Because we excluded a mutual interference of ATP and nucleosome binding, we postulate that both motifs stimulate the ATPase and chromatin-remodeling activities upon binding of BRG1 to nucleosomes, probably via allosteric mechanisms. Furthermore, mutations of both motifs similarly affect the enzymatic functionality of BRG1 in vitro and in living cells. Of note, in BRG1deficient H1299 cells, exogenously expressed wtBRG1, but not BRG1 Q758A and BRG1 K785R, exhibited a tumor suppressorlike function.

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“…To delete the floxed allele in primordial germ cells (embryonic day 15.5, Fig. 2C) (18,19), male Ddx4-Cre; Chd4 WT/Δ were crossed with Chd4 fl/fl females to generate Ddx4-Cre;…”

Section: Deletion Of Chd4 But Not Chd3 Results In Testis Developmentamentioning

confidence: 99%

Mouse CHD4-NURD is required for neonatal spermatogonia survival and normal gonad development

Castro

Onate-Colobon

Previato

et al. 2019

Preprint

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Testis development and sustained germ cell production in adults rely on the establishment of spermatogonia stem cells and their proper differentiation into mature gametes. Control of these processes involves not only promoting the expression of genes required for cell 2 survival and differentiation but also repressing other cell fates. This level of transcriptional control requires chromatin-remodeling complexes that restrict or promote transcription machinery. Here, we investigated the roles of the NUcleosome Remodeling and Deacetylase (NURD) complex during spermatogenesis. Our cytological and biochemical analyses revealed differential expression and composition of NURD subunits in gametocytes at different stages of testis development. Germ cell-specific deletion of the NURD catalytic component CHD4, but not CHD3, resulted in male infertility due to failed maintenance of the undifferentiated spermatogonia stem cell population. Genome-wide CHD4 localization and transcriptomic analyses revealed that CHD4 binds the promoters and regulates the expression of genes involved in spermatogonia cell survival, and differentiation. These results uncover the requirements of CHD4 in mammalian gonad development, and point to unique roles for the NURD complex with respect to other chromatin remodelers during gamete development. Significance StatementGametogenesis is a fundamental developmental program required for sustained fertility and survival of all sexually reproducing species. The developing male gamete undergoes numerous cell divisions and developmental stage transitions that are carefully monitored by epigenetic mechanisms. One prominent mechanism is directed by chromatin remodeling complexes, which modify chromatin structure and thereby control fundamental cellular processes such as gene transcription. In this work, we focused in understanding the role of CHD4 and CHD3 proteins, catalytic subunits of the NURD chromatinremodeling complex, in mouse gametogenesis. We find that CHD4 has an essential function in gametogenesis, with an absolute requirement for maintenance of and differentiation of spermatogonia populations in the developing testis. This is achieved by CHD4-mediated transcriptional regulation of genes important for spermatogonia survival, and differentiation.

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CHD3 and CHD4 form distinct NuRD complexes with different yet overlapping functionality

Cited by 76 publications

References 119 publications

Maintenance of Genome Integrity by Mi2 Homologs CHD-3 and LET-418 in Caenorhabditis elegans

Maintenance of Genome Integrity by Mi2 Homologs CHD-3 and LET-418 in Caenorhabditis elegans

Elucidation of the functional roles of the Q and I motifs in the human chromatin-remodeling enzyme BRG1

Mouse CHD4-NURD is required for neonatal spermatogonia survival and normal gonad development

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