Exploring the Molecular Underpinnings of Cancer-Causing Oncohistone Mutants Using Yeast as a Model.

Understanding the molecular basis of cancer initiation and progression is critical in developing effective treatment strategies. Recently, mutations in genes encoding histone proteins that drive oncogenesis have been identified, converting these essential proteins into "oncohistones". Understanding how oncohistone mutants, which are commonly single missense mutations, subvert the normal function of histones to drive oncogenesis requires defining the functional consequences of such changes. Histones genes are present in multiple copies in the human genome with 15 genes encoding histone H3 isoforms, the histone for which the majority of oncohistone variants have been analyzed thus far. With so many wildtype histone proteins being expressed simultaneously within the oncohistone, it can be difficult to decipher the precise mechanistic consequences of the mutant protein. In contrast to humans, budding and fission yeast contain only two or three histone H3 genes, respectively. Furthermore, yeast histones share ~90% sequence identity with human H3 protein. Its genetic simplicity and evolutionary conservation make yeast an excellent model for characterizing oncohistones. The power of genetic approaches can also be exploited in yeast models to define cellular signaling pathways that could serve as actionable therapeutic targets. In this review, we focus on the value of yeast models to serve as a discovery tool that can provide mechanistic insights and inform subsequent translational studies in humans.

Hyperextended telomeres promote formation of C-circle DNA in telomerase positive human cells.

Telomere length maintenance is crucial to cancer cell immortality. Up to 15% of cancers utilize a telomerase-independent, recombination-based mechanism termed alternative lengthening of telomeres (ALT). Currently, the primary ALT biomarker is the C-circle, a type of circular DNA with extrachromosomal telomere repeats (cECTRs). How C-circles form is not well characterized. We investigated C-circle formation in the human cen3tel cell line, a long-telomere, telomerase+ (LTT+) cell line with progressively hyper-elongated telomeres (up to ∼100 kb). cECTR signal was observed in 2D gels and C-circle assays but not t-circle assays, which also detect circular DNA with extrachromosomal telomere repeats. Telomerase activity and C-circle signal were not separable in the analysis of clonal populations, consistent with C-circle production occurring within telomerase+ cells. We observed similar cECTR results in two other LTT+ cell lines, HeLa1.3 (∼23 kb telomeres) and HeLaE1 (∼50 kb telomeres). In LTT+ cells, telomerase activity did not directly impact C-circle signal; instead, C-circle signal correlated with telomere length. LTT+ cell lines were less sensitive to hydroxyurea than ALT+ cell lines, suggesting that ALT status is a stronger contributor to replication stress levels than telomere length. Additionally, the DNA repair-associated protein FANCM did not suppress C-circles in LTT+ cells as it does in ALT+ cells. Thus, C-circle formation may be driven by telomere length, independently of telomerase and replication stress, highlighting limitations of C-circles as a stand-alone ALT biomarker.

pH-equilibration of human hair: Kinetics and pH-dependence of the partition ratios for H+ - and OH- -ions based on a Freundlich isotherm.

Hair is an insoluble, fibrous, α-keratinous, protein composite material, providing outer coverage, e.g., for mammals. In the context of a wider study on the effects of pH on human hair properties, we investigated the time-dependence of pH-equilibration study across the acid and the basic pH-range, using appropriate pure solutions of hydrochloric acid and sodium hydroxide. The results show that pH-equilibration follows essentially equal 1st-order kinetics across the pH-range. The characteristic process time does not change significantly and is in the range of 2.5-5 h. The analysis enables to determine the equilibrium uptakes of H+- and OH- -ions. These follow the expected U-shaped path across the pH-range. For both acidic and alkaline conditions, data are well described by two very similar sorption isotherms of the Freundlich-type. In consequence, partition ratios for both ions are highest near neutrality (pH 7: >6000) and drop off strongly towards low and high pHs (<50). Hair is thus a very strong 'sink' for H+ and OH-. This observation fundamentally challenges traditional views of limited ion uptake, namely, in the mid-pH-range due to hindered diffusion. It also does not support considerations on special roles of certain pHs, specific groups of amino acids, or morphological components. Our analysis thus suggests that established views of the interaction of hair and pH need to be reconsidered, The Freundlich isotherm approach appears to provide a versatile tool to refine our understanding of the interactions of hair and possibly other keratinous materials (horn, nail, feathers) with acids and bases.

Hyperextended telomeres promote C-circle formation in telomerase positive human cells.

Telomere length maintenance is crucial to cancer cell immortality. Up to 15% of cancers utilize a telomerase-independent, recombination-based mechanism termed alternative lengthening of telomeres (ALT). The primary ALT biomarker is the C-circle, a type of circular DNA with extrachromosomal telomere repeats (cECTRs). How C-circles form is not well characterized. To investigate C-circle formation in telomerase+ cells, we studied the human cen3tel cell line, in which telomeres progressively hyper-elongated post TERT -immortalization. cECTR signal was observed in 2D gels and C-circle assays but not t-circle assays, which also detect cECTRs. Telomerase activity and C-circle signal were not separable in the analysis of clonal populations, consistent with C-circle production occurring within telomerase+ cells. Two other long telomere, telomerase+ (LTT+) cell lines, HeLa1.3 (~23 kb telomeres) and HeLaE1 (~50 kb telomeres), had similar cECTR properties. Telomerase activity did not directly impact C-circle signal in LTT+ cells; instead, C-circle signal correlated with telomere length. LTT+ lines were less sensitive to hydroxyurea than an ALT+ cell line, suggesting that ALT status is a stronger contributor to replication stress levels than telomere length. Additionally, FANCM did not suppress C-circles in LTT+ cells as it does in ALT+ cells. Thus, C-circle formation may be driven by telomere length, independently of telomerase and replication stress, highlighting limitations of C-circles as a stand-alone ALT biomarker.

Perm-waved human hair: a thermorheologically complex shape memory composite.

A "permanent" bent shape can be imposed on a straight human hair by a two-stage reduction/oxidation (perm-waving) process. The process relies on the molecular level on sulfhydryl/disulfide interchange as bond exchange reaction (BER). We expected a well-documented transition temperature around 60°C to be the trigger for the shape memory (SM) process of perm-waved hair. We confirm the existence of the SM process as such and investigate its time and temperature dependence. The results show a two-stage SM behavior, implying two distinct variations of the BER. The model to fit the data contains two fractional, normalized, elastic bending rigidities, which are strictly compensatory. They show Arrhenius-type temperature dependence and a common activation energy (EA) of ∼-12 kJ/mol. The characteristic relaxation time for the first SM process shows little, if any, temperature dependence (EA = -4 ± 2.7 kJ/mol). This is in contrast to the second process (EA = -58 ± 5.5 kJ/mol) but in line with the expected properties of the suggested BERs. None of the parameters shows any sign of the expected trigger transition (∼60°C). We hypothesize that this specific transition occurs only for large tensile deformations, when specific SS bonds in the intermediate filaments of hair are activated. There is thus no specific "trigger" transition for the SM behavior of bent, perm-waved hair.

Textile materials inspired by structural colour in nature.

The concept of mimicking structural colour in nature as an alternative to traditional textile coloration techniques would reduce dependency on dyes, pigments and vast quantities of water in the textile supply chain. Structural colours originate from the physical interaction of light with nanoscale structures. This is exhibited in the bodies and wings of certain species of butterfly, beetles and plants. The angular optical effects of the Chrysina gloriosa beetle result from the periodicity due to the cholesteric liquid crystal (CLC) structure adopted by the cells in their exoskeleton. The optical properties of CLCs makes promising applications for optical sensors and anti-counterfeit materials. Application using inkjet printing technology enables designs to be tuned to meet product requirements, and with a hydrophobic treatment challenges associated with a rough surface such as textiles are overcome. Here we report inkjet printing CLC solutions onto hydrophobic pre-treated textiles. CIE L*a*b* values demonstrate the resultant colourful films display a greater degree of colour compared to those on untreated textiles.