title: 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.
authors.affiliation: Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA Driskill Graduate Program in Life Sciences, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.
authors.forename: Aurimas
authors.identifier: http://orcid.org/0000-0003-0832-8302
authors.initials: A
authors.lastname: Vinckevicius
authors.affiliation: Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA.
authors.forename: J Brandon
authors.initials: JB
authors.lastname: Parker
authors.affiliation: Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA Driskill Graduate Program in Life Sciences, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA Department of Pharmacology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA Robert H. Lurie Comprehensive Cancer Center, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA debu@northwestern.edu.
authors.forename: Debabrata
authors.initials: D
authors.lastname: Chakravarti
mesh_terms: D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002843:Chromatin / Q000235:genetics; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004268:DNA-Binding Proteins / Q000235:genetics; D015870:Gene Expression; D057809:HEK293 Cells; D006367:HeLa Cells; D051863:Host Cell Factor C1 / Q000235:genetics*; D006801:Humans; D046912:Multiprotein Complexes / Q000235:genetics*; D059372:Nucleotide Motifs / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D017393:RNA Editing / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D051863:Host Cell Factor C1; D046912:Multiprotein Complexes; D012097:Repressor Proteins; C498470:THAP11 protein, human; D015534:Trans-Activators; C114253:ZNF143 protein, human