SUMMARY Upon androgen stimulation, PKN1-mediated histone H3 threonine 11 phosphorylation (H3T11P) promotes AR target genes activation. However, the underlying mechanism is not completely understood. Here, we show that WDR5, a subunit of the SET1/MLL complex, interacts with H3T11P and this interaction facilitates the recruitment of the MLL1 complex and subsequent H3K4 trimethylation (H3K4me3). Using ChIP-seq, we find that androgen stimulation results in a six-fold increase in the number of H3T11P-marked regions and induces WDR5 colocalization to one third of H3T11P-enriched promoters, thus establishing a genome-wide relationship between H3T11P and recruitment of WDR5. Accordingly, PKN1 knock-down or chemical inhibition severely blocks WDR5 association and H3K4me3 on AR target genes. Finally, WDR5 is critical in prostate cancer cell proliferation, and is hyperexpressed in human prostate cancers. Together, these results identify WDR5 as a critical epigenomic integrator of histone phosphorylation and methylation and a major driver of androgen-dependent prostate cancer cell proliferation.

Summary Host cell factor-1 (HCF-1) is a metazoan transcriptional co-regulator essential for cell cycle progression and cell proliferation. Current models suggest a mechanism whereby HCF-1 functions as a direct co-regulator of E2F proteins, facilitating the expression of genes necessary for cell proliferation. In this report, we show that HCF-1 recruitment to numerous E2F-bound promoters is mediated by the concerted action of zinc finger transcription factors THAP11 and ZNF143, rather than E2F proteins directly. THAP11, ZNF143, and HCF-1 form a mutually dependent complex on chromatin, which is independent of E2F occupancy. Disruption of the THAP11/ZNF143/HCF-1 complex results in altered expression of cell cycle control genes and leads to reduced cell proliferation, cell cycle progression, and cell viability. These data establish a new model which suggests that a THAP11/ZNF143/HCF-1 complex is a critical component of the transcriptional regulatory network governing cell proliferation.

In the article by Kim et al., we the authors made some errors during final preparation of a few figures (mostly on proper marking of lanes). We have corrected these errors as follows:Under Figures 1A, 1D, and S1F, we failed to indicate that the adjacent lanes are non-continuous but from the same gel. Lines have now been added to indicate this.In Figure 1C and its corresponding Figure S1F, under Coomassie blue, we inadvertently ran and presented four or three, respectively, lanes of purified GST-WDR5 input. We can see how this could be confusing, so now we are only showing one lane. As also noted in the original paper, input lanes are identical in Figures 1C and S1F. These corrections do not alter the conclusions of the original paper; however, we apologize for any inconvenience or confusion that this may have caused. Figure 1. Revised Panels from Original Figure 1 and Figure S1 Molecular Cell 58, 557, May 7, 2015 ª2015 Elsevier Inc. 557

The THAP11 and ZNF143 transcription factors recognize overlapping DNA sequences and are reported to exhibit signs of both competitive and cooperative binding. HCFC1 serves as a scaffold protein, bridging interactions between transcription factors, including THAP11 and ZNF143, and transcriptional coregulators. The exact mechanism of how DNA sequences guide the recruitment of the THAP11/ZNF143/HCFC1 complex to chromatin is still controversial. In this study, we use chromosomally integrated synthetic constructs and clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9-mediated approaches in intact cells to elucidate the role of the DNA sequence in the recruitment of this complex and to establish its biological relevance. We show that the ACTACA submotif, shared by both THAP11 and ZNF143, directs the recruitment of THAP11 and HCFC1 to ZNF143-occupied loci. Importantly, its position, spacing, and orientation relative to the ZNF143 core motif are critical for this action. CRISPR-Cas9-mediated alterations of the ACTACA submotif at endogenous promoters recapitulated results obtained with synthetic constructs and resulted in altered gene transcription and histone modifications at targeted promoters. Our in vivo approaches provide strong evidence for the molecular role of the ACTACA submotif in THAP11, ZNF143, and HCFC1 cooperative recruitment to chromatin and its biological role in target gene expression. Host cell factor 1 (HCFC1) is an atypical transcriptional coregulator that is translated as a single 2,035-amino-acid peptide and undergoes proteolytic cleavage at the centrally located PRO repeats (1, 2), and the resulting N and C termini noncovalently reassociate via two pairs of self-association sequences (3, 4). The N-terminal fragment of HCFC1 contains a six-Kelch-repeat ␤-propeller (Kelch domain) and a basic region, both of which facilitate protein-protein association. The Kelch domain recognizes a 4-amino-acid ([E/D]HXY) HCFC1 binding motif (HBM) (5) found in a large number of transcription factors and cofactors (6), including LZIP (5), Set1 (7), E2F4 (8), and the THAP family of proteins (9). Recently reported findings suggest that HCFC1 also associates with ZNF143 via its Kelch domain (10), but the mechanism of this interaction remains unclear, because ZNF143 lacks the HBM. The basic region of HCFC1 mediates associations between a distinct set of proteins, such as GABP (11), Sin3 (7), and Sp1 (12), but can also bind proteins associated with the Kelch domain, as exemplified by E2F4 (8).HCFC1 is conserved in metazoans and has been implicated in playing critical roles in cell cycle regulation and proliferation (13-18). A single-point mutation in the HCFC1 Kelch domain in the temperature-sensitive hamster cell line tsBN67 causes HCFC1 dissociation from chromatin at a nonpermissive temperature, leading to cell cycle arrest in G 1 phase as well as defects in cytokinesis (14,17,18). Similar cell cycle aberrations were noted for HeLa cells upon small interfering RNA (siRNA)-mediated knockdown of HCFC1 (...

Recent decades have been filled with groundbreaking research in the field of endocrine hormone signaling. Pivotal events like the isolation and purification of the estrogen receptor, the cloning of glucocorticoid receptor cDNA, or dissemination of nuclear hormone receptor (NHR) DNA binding sequences are well recognized for their contributions. However, the novel genome-wide and gene-specific information obtained over the last decade describing NHR association with chromatin, cofactors, and epigenetic modifications, as well as their role in gene regulation, has been largely facilitated by the adaptation of the chromatin immunoprecipitation (ChIP) technique. Use of ChIP-based technologies has taken the field of hormone signaling from speculating about the transcription-enabling properties of acetylated chromatin and putative transcription (co-)factor genomic occupancy to demonstrating the detailed, stepwise mechanisms of factor binding and transcriptional initiation; from treating hormone-induced transcription as a steady-state event to understanding its dynamic and cyclic nature; from looking at the DNA sequences recognized by various DNA-binding domains in vitro to analyzing the cell-specific genome-wide pattern of nuclear receptor binding and interpreting its physiological implications. Not only have these events propelled hormone research, but, as some of the pioneering studies, have also contributed tremendously to the field of molecular endocrinology as a whole. In this review, we give a brief summary of some of the most important discoveries in hormone signaling using ChIP and other derivative techniques and speculate on what the future may hold.