Genomic Determinants of THAP11/ZNF143/HCFC1 Complex Recruitment to Chromatin

Abstract

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.

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Supplemental material for this article may be found at http://dx.doi.org/10.1128/MCB.00477-15.

ACKNOWLEDGMENTS

Flow cytometry work was performed at the Robert H. Lurie Comprehensive Cancer Center of Northwestern University Flow Cytometry Core Facility, which is supported by Cancer Center support grant NCI CA060553. This work was supported in part by NCI grant R01 CA133755 (D.C.), the Robert H. Lurie Comprehensive Cancer Center, and the Department of Obstetrics and Gynecology at Northwestern University.

A.V., J.B.P., and D.C. conceived the research plan, designed experiments, and analyzed data. A.V. performed research and wrote the draft manuscript. All authors reviewed and edited the manuscript.

We declare that we have no competing financial interests.