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In eukaryotes, DNA is compacted into a complex structure known as chromatin. The unravelling of DNA is a crucial step in DNA repair, replication, transcription and recombination as this allows access to DNA for these processes. Failure to package DNA into the nucleosome, the individual unit of chromatin, can lead to genomic instability, driving a cell into apoptosis, senescence, or cellular proliferation. Ultraviolet (UV) radiation damage causes destabilisation of chromatin integrity. UV irradiation induces DNA damage such as photolesions and subjects the chromatin to substantial rearrangements, causing the arrest of transcription forks and cell cycle arrest. Highly conserved processes known as nucleotide and base excision repair (NER and BER) then begin to repair these lesions. However, if DNA repair fails, the cell may be forced into apoptosis. The modification of various histones as well as nucleosome remodelling via ATP-dependent chromatin remodelling complexes are required not only to repair these UV-induced DNA lesions, but also for apoptosis signalling. Histone modifications and nucleosome remodelling in response to UV also lead to the recruitment of various repair and pro-apoptotic proteins. Thus, the way in which a cell responds to UV irradiation via these modifications is important in determining its fate. Failure of these DNA damage response steps can lead to cellular proliferation and oncogenic development, causing skin cancer, hence these chromatin changes are critical for a proper response to UV-induced injury.

Ultraviolet radiation (UV) from sunlight is the primary cause of skin and ocular neoplasia. Brahma (BRM) is part of the SWI/SNF chromatin remodeling complex. It provides energy for rearrangement of chromatin structure. Previously we have found that human skin tumours have a hotspot mutation in BRM and that protein levels are substantially reduced. Brm−/− mice have enhanced susceptibility to photocarcinogenesis. In these experiments, Brm−/− mice, with both or a single Trp53 allele were exposed to UV for 2 or 25 weeks. In wild type mice the central cornea and stroma became atrophic with increasing time of exposure while the peripheral regions became hyperplastic, presumably as a reparative process. Brm−/−, Trp53+/−, and particularly the Brm−/− Trp53+/− mice had an exaggerated hyperplastic regeneration response in the corneal epithelium and stroma so that the central epithelial atrophy or stromal loss was reduced. UV induced hyperplasia of the epidermis and corneal epithelium, with an increase in the number of dividing cells as determined by Ki-67 expression. This response was considerably greater in both the Brm−/− Trp53+/+ and Brm−/− Trp53+/− mice indicating that Brm protects from UV-induced enhancement of cell division, even with loss of one Trp53 allele. Cell division was disorganized in Brm−/− mice. Rather than being restricted to the basement membrane region, dividing cells were also present in the suprabasal regions of both tissues. Brm appears to be a tumour suppressor gene that protects from skin and ocular photocarcinogenesis. These studies indicate that Brm protects from UV-induced hyperplastic growth in both cutaneous and corneal keratinocytes, which may contribute to the ability of Brm to protect from photocarcinogenesis.

The phenotypic expression of quantitative characters is a function of the individual's genotype and the environment in which it is measured. In a previous reciprocal transplant study, we found that patterns of genetic differences in resistance to herbivores among adjacent subpopulations of northern red oak (Quercus rubra L.), were consistent with a local adaptation hypothesis. The goal of this study was to determine if variation in water availability may have been a mechanism responsible for these previously observed patterns. In 1989 a common garden study was initiated using acorns from maternal trees occupying either a north- or south-facing slope microhabitat in an oak-hickory forest in east central Missouri, USA. The seedlings were grown under one of two water treatments, irrigated or natural. In 1992, we utilized this experiment to examine the quantitative character of the percentage of leaf area damaged by herbivores, which is a measure of the phenotypically expressed level of resistance. Specifically, we made three predictions: (1) because northern red oak seem to grow best in mesic environments, seedlings receiving more water should show greater resistance to herbivores; (2) if the subpopulations from north- and south-facing slope microhabitats are genetically differentiated with respect to the quantitative character of resistance to herbivores, then there will be a significant effect of maternal slope microhabitat on the percentage of leaf area damaged; and (3) if the pattern of resistance to herbivores found among subpopulations reflects local adaptation to moisture levels in their own microhabitat, then we would expect to find a significant maternal slope microhabitat by water treatment interaction, with north-facing slope seedlings incurring less damage in the wetter (irrigated) treatment and south-facing slope seedlings incurring less damage in the drier (natural) treatment. Our data supported the first two predictions: seedlings in the irrigated treatment showed a significantly lower percentage of leaf area damage than those in the natural treatment, and the percentage of leaf area damaged was significantly lower on seedlings from maternal plants occupying the north-facing slope microhabitat. However, we found no significant interaction between maternal slope microhabitat and water treatment. These findings demonstrate that northern red oak supbopulations respond phenotypically to water availability, but this factor does not appear to be the underlying mechanism behind the previously observed local adaptation expressed as resistance to herbivores.

Aulacoseira ambigua abundance and filament length were measured weekly during spring and autumn bloom periods in Trout Lake, Wisconsin, USA. In addition, several chemical and biological variables thought to influence A. ambigua growth were assessed. Results of the field-based observations were complimented with controlled laboratory experiments to evaluate the effects of phosphorus availability on A. ambigua growth. Relative to the autumn bloom period, A. ambigua colonies were generally larger and more abundant in spring prior to the termination of the bloom in June. Chlorophyll-a concentrations indicated that other phytoplankton were also more abundant during the spring bloom. Batch culture experiments indicated that increased phosphorus availability during the spring bloom contributed to the seasonal increase in A. ambigua filament length and abundance. Increase in A. ambigua filament length in response to increased phosphorus availability is discussed as a mechanism that may increase nutrient removal through sedimentation and subsequently decrease the efficiency of nutrient regeneration in higher productivity lakes.

Oncogenic mutations in the small GTPase RAS contribute to ~30% of human cancers. In a Drosophila genetic screen, we identified novel and evolutionary conserved cancer genes that affect Ras-driven tumorigenesis and metastasis in Drosophila including confirmation of the tetraspanin Tsp29Fb. However, it was not known whether the mammalian Tsp29Fb orthologue, TSPAN6, has any role in RAS-driven human epithelial tumors. Here we show that TSPAN6 suppressed tumor growth and metastatic dissemination of human RAS activating mutant pancreatic cancer xenografts. Whole-body knockout as well as tumor cell autonomous inactivation using floxed alleles of Tspan6 in mice enhanced KrasG12D-driven lung tumor initiation and malignant progression. Mechanistically, TSPAN6 binds to the EGFR and blocks EGFR-induced RAS activation. Moreover, we show that inactivation of TSPAN6 induces an epithelial-to-mesenchymal transition and inhibits cell migration in vitro and in vivo. Finally, low TSPAN6 expression correlates with poor prognosis of patients with lung and pancreatic cancers with mesenchymal morphology. Our results uncover TSPAN6 as a novel tumor suppressor receptor that controls epithelial cell identify and restrains RAS-driven epithelial cancer.

Background: Ultraviolet radiation (UVR) is the principal cause of keratinocyte skin cancers. Previous work found that levels of the chromatin remodelling protein, Brahma (Brm), are diminished during the progression from actinic keratoses to cutaneous squamous cell carcinomas in humans, and its loss in UVirradiated mouse skin causes epidermal hyperplasia and increased tumour incidence. Methods: The skins of mice and mouse and human keratinocytes deficient in Brm were exposed to UVR and evaluated for cell cycle progression and DNA damage response.Objective: To identify the mechanisms by which loss of Brm contributes to UVR-induced skin carcinogenesis. Results: In both mouse keratinocytes and HaCaT cells, Brm deficiency led to an increased cell population growth following UVR exposure compared to cells with normal levels of Brm. Cell cycle analysis using a novel assay showed that Brm-deficient keratinocytes entered cell cycle arrest normally, but escaped from cell cycle arrest faster, enabling them to begin proliferating earlier. In mouse keratinocytes, Brm primarily affected accumulation in G 0 /G 1 -phase, whereas in HaCaT cells, which lack normal p53, accumulation in G 2 /M-phase was affected. Brm-deficient keratinocytes in mouse skin and human cell cultures also had higher levels of UVR-induced cyclobutane pyrimidine dimer photolesions. These effects occurred without any compensatory increase in DNA repair or cell death to remove photolesions or the cells that harbor them from the keratinocyte population. Conclusion:The loss of Brm in keratinocytes exposed to UVR enables them to resume proliferation while harboring DNA photolesions, leading to an increased fixation of mutations and, consequently, increased carcinogenesis.

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