Na+ and K+ Ions Differently Affect Nucleosome Structure, Stability, and Interactions with Proteins

Inorganic ions are essential factors stabilizing nucleosome structure; however, many aspects of their effects on DNA transactions in chromatin remain unknown. Here, differential effects of K+ and Na+ on the nucleosome structure, stability, and interactions with protein complex FACT (FAcilitates Chromatin Transcription), poly(ADP-ribose) polymerase 1, and RNA polymerase II were studied using primarily single-particle Förster resonance energy transfer microscopy. The maximal stabilizing effect of K+ on a nucleosome structure was observed at ca. 80–150 mM, and it decreased slightly at 40 mM and considerably at >300 mM. The stabilizing effect of Na+ is noticeably lower than that of K+ and progressively decreases at ion concentrations higher than 40 mM. At 150 mM, Na+ ions support more efficient reorganization of nucleosome structure by poly(ADP-ribose) polymerase 1 and ATP-independent uncoiling of nucleosomal DNA by FACT as compared with K+ ions. In contrast, transcription through a nucleosome is nearly insensitive to K+ or Na+ environment. Taken together, the data indicate that K+ environment is more preserving for chromatin structure during various nucleosome transactions than Na+ environment.

Variations in Nucleosome Structure and Organization among Eukaryotes

Folding eukaryotic DNA by chromatin is a vital process necessary for the proper function of DNA. This is achieved by the fundamental unit of chromatin, known as a nucleosome. The position of a nucleosome and its interaction with DNA plays a crucial role in regulating the vital processes involved in DNA function. Factors such as variations in nucleosome and its core structure and histone fold variations will help to understand nucleosome functions and their role in DNA replication, transcription, translation, posttranslational modifications, re-combinations and repair. The present review focuses on recent findings in understanding the variations in the structure and functions of nucleosomes across eukaryotes. Variations in the nucleosome organization and its assembly have also been discussed by stating the contribution of histone binding factors and chromatin assembly factors.

Histone sequence variation in divergent eukaryotes facilitates diversity in chromatin packaging

The histone proteins defining nucleosome structure are highly conserved in common model organisms and are frequently portrayed as uniform chromatin building blocks. We surveyed over 1700 complete eukaryotic genomes and confirm that almost all encode recognisable canonical core histones. Nevertheless, divergent eukaryotes show unrecognised diversity in histone sequences and offer an opportunity to observe the potential for nucleosome variation. Recombinant histones for Plasmodium falciparum, Giardia lamblia, Encephalitozoon cuniculi and Leishmania major were prepared alongside those for human, Xenopus laevis and Saccharomyces cerevisiae. All could be assembled into nucleosomes in vitro on sequences known to direct positioning with metazoan histones. P. falciparum histones refolded into very stable nucleosomes consistent with a highly regulated transcriptional programme. In contrast, G. lamblia and E. cuniculi histones formed less stable nucleosomes and were prone to aggregation as H3-H4 tetramers. Inspection of the histone fold dimer interface residues suggested a potential to form tetrasomal arrays consistent with polymerisation. DNA binding preferences observed using systematic evolution of ligands by exponential enrichment (SELEX) for human, P. falciparum and E. cuniculi histone octamers were highly similar and reflect a shared capability to package diverse genomic sequences. This demonstrates that nucleosomal organisation is retained across eukaryotes and can accommodate genome variation, but histone protein sequences vary more than commonly recognised to provide the potential for diversity of chromatin features.

Histone Variants: The Nexus of Developmental Decisions and Epigenetic Memory

Nucleosome dynamics and properties are central to all forms of genomic activities. Among the core histones, H3 variants play a pivotal role in modulating nucleosome structure and function. Here, we focus on the impact of H3 variants on various facets of development. The deposition of the replicative H3 variant following DNA replication is essential for the transmission of the epigenomic information encoded in posttranscriptional modifications. Through this process, replicative H3 maintains cell fate while, in contrast, the replacement H3.3 variant opposes cell differentiation during early embryogenesis. In later steps of development, H3.3 and specialized H3 variants are emerging as new, important regulators of terminal cell differentiation, including neurons and gametes. The specific pathways that regulate the dynamics of the deposition of H3.3 are paramount during reprogramming events that drive zygotic activation and the initiation of a new cycle of development.

Genome-wide Identification and Functional Analysis of the H3 Gene Family in Cotton

Background: Histones are major components of chromatin, which is a nucleosome structure associated with chromosome segregation, DNA packaging and transcriptional regulation. Histone H3 is encoded by many genes in most eukaryotic species, but little information is known about the Histone H3 gene family in cotton.Results: In this study, we identified and analyzed the evolution and expression of histone H3 gene family in cotton. First, 34 G. hirsutum genes were identified belonging to the H3 gene family which were divided into four subclasses: CENH3, H3.1, H3.3 and H3-like. Among these H3.1 subclass contained the highest number of genes (22 members) followed by H3.3 subclass (9 members). In addition, there were18 and 16 H3 genes identified in G. arboretum and G. raimondii, respectively. Furthermore, we conducted conserved sequence analysis of H3 proteins, and found that the four amino acids signature including A31F41S87A90 for H3.1 and T31Y41H87L90 for H3.3 could be used to discriminate H3.1 from H3.3. The expression of H3 gene family varied in different tissues and developmental stages of G. hirsutum, where H3.1 subclass genes play a critical role in pistil development. By virus-induced gene silencing of GhCENH3 (Gh_D07G1382) gene, the size of leaf got smaller with pYL156-CENH3 than that with pYL156 in TM-1. Whereas, the number of the stomata in the leaf epidermis and number of chloroplasts in the leaf stomatal guard cells by pYL156-CENH3 was more than that by pYL156 and pYL156-PDS.Conclusions: Four sub-classes (CENH3, H3.1, H3.3 and H3-like) of H3 gene family were highly conserved in cotton during the rapid phase of evolution among which CENH3 is necessary for leaf growth. These findings are useful for providing further insights into cotton biology and breeding.

Regulation of the Mammalian SWI/SNF Family of Chromatin Remodeling Enzymes by Phosphorylation during Myogenesis

Myogenesis is the biological process by which skeletal muscle tissue forms. Regulation of myogenesis involves a variety of conventional, epigenetic, and epigenomic mechanisms that control chromatin remodeling, DNA methylation, histone modification, and activation of transcription factors. Chromatin remodeling enzymes utilize ATP hydrolysis to alter nucleosome structure and/or positioning. The mammalian SWItch/Sucrose Non-Fermentable (mSWI/SNF) family of chromatin remodeling enzymes is essential for myogenesis. Here we review diverse and novel mechanisms of regulation of mSWI/SNF enzymes by kinases and phosphatases. The integration of classic signaling pathways with chromatin remodeling enzyme function impacts myoblast viability and proliferation as well as differentiation. Regulated processes include the assembly of the mSWI/SNF enzyme complex, choice of subunits to be incorporated into the complex, and sub-nuclear localization of enzyme subunits. Together these processes influence the chromatin remodeling and gene expression events that control myoblast function and the induction of tissue-specific genes during differentiation.