General regulatory factors exert differential effects on nucleosome sliding activity of the ISW1a complex.
Andrea Oyarz��n-Cisterna, Cristi��n Gidi, Fernanda Raiqueo, Roberto Amigo, Camila Rivas, Marcela Torrej��n, Jos�� L Guti��rrez
Author Information
Andrea Oyarz��n-Cisterna: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Cristi��n Gidi: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Fernanda Raiqueo: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Roberto Amigo: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Camila Rivas: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Marcela Torrej��n: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile.
Jos�� L Guti��rrez: Departamento de Bioqu��mica y Biolog��a Molecular, Facultad de Ciencias Biol��gicas, Universidad de Concepci��n, 4070043, Concepci��n, Chile. lgutier@udec.cl. ORCID
BACKGROUND: Chromatin dynamics is deeply involved in processes that require access to DNA, such as transcriptional regulation. Among the factors involved in chromatin dynamics at gene regulatory regions are general regulatory factors (GRFs). These factors contribute to establishment and maintenance of nucleosome-depleted regions (NDRs). These regions are populated by nucleosomes through histone deposition and nucleosome sliding, the latter catalyzed by a number of ATP-dependent chromatin remodeling complexes, including ISW1a. It has been observed that GRFs can act as barriers against nucleosome sliding towards NDRs. However, the relative ability of the different GRFs to hinder sliding activity is currently unknown. RESULTS: Considering this, we performed a comparative analysis for the main GRFs, with focus in their ability to modulate nucleosome sliding mediated by ISW1a. Among the GRFs tested in nucleosome remodeling assays, Rap1 was the only factor displaying the ability to hinder the activity of ISW1a. This effect requires location of the Rap1 cognate sequence on linker that becomes entry DNA in the nucleosome remodeling process. In addition, Rap1 was able to hinder nucleosome assembly in octamer transfer assays. Concurrently, Rap1 displayed the highest affinity for and longest dwell time from its target sequence, compared to the other GRFs tested. Consistently, through bioinformatics analyses of publicly available genome-wide data, we found that nucleosome occupancy and histone deposition in vivo are inversely correlated with the affinity of Rap1 for its target sequences in the genome. CONCLUSIONS: Our findings point to DNA binding affinity, residence time and location at particular translational positions relative to the nucleosome core as the key features of GRFs underlying their roles played in nucleosome sliding and assembly.
