Chronic Replication Problems Impact Cell Morphology and Adhesion of DNA Ligase I Defective Cells

Moderate DNA damage resulting from metabolic activities or sub-lethal doses of exogenous insults may eventually lead to cancer onset. Human 46BR.1G1 cells bear a mutation in replicative DNA ligase I (LigI) which results in low levels of replication-dependent DNA damage. This replication stress elicits a constitutive phosphorylation of the ataxia telangiectasia mutated (ATM) checkpoint kinase that fails to arrest cell cycle progression or to activate apoptosis or cell senescence. Stable transfection of wild type LigI, as in 7A3 cells, prevents DNA damage and ATM activation. Here we show that parental 46BR.1G1 and 7A3 cells differ in important features such as cell morphology, adhesion and migration. Comparison of gene expression profiles in the two cell lines detects Bio-Functional categories consistent with the morphological and migration properties of LigI deficient cells. Interestingly, ATM inhibition makes 46BR.1G1 more similar to 7A3 cells for what concerns morphology, adhesion and expression of cell-cell adhesion receptors. These observations extend the influence of the DNA damage response checkpoint pathways and unveil a role for ATM kinase activity in modulating cell biology parameters relevant to cancer progression

Assisted-selection of naturally caffeine-free coffee cultivars-characterization of SNPs from a methyltransferase gene.

Breeding of caffeine-free coffee cultivars require tools for an early selection of progenies bearing this later trait. Genes from caffeine synthesis and degradation represent major targets for the development of molecular markers for assisted selection. In this study, we characterized SNPs identified on the caffeine synthase gene from AC1 mutant, a naturally caffeine-free arabica coffee plant. Molecular analysis of normal and mutant sequences indicates the occurrence of SNPs in protein domains, potentially associated with caffeine synthesis in coffee. Progenies F2, F1BC1 and BC from crosses of AC mutants and elite cultivars were evaluated regarding caffeine content in grains and genomic segregation profile of selected SNPs. Genotyping analysis allowed the discrimination between homozygous and heterozygous plants. Quantification of caffeine content indicated a significant variability among progenies and a low frequency of caffeine-free plants. Statistical analyses of genotyping and phenotyping results showed significant association between presence of selected SNPs and reduced caffeine content. Moreover, this association occurs through all evaluated genetic backgrounds and generations, indicating an inheritance stability of both trait and markers. The molecular markers described here represent a successful case of assistedselection in coffee, indicating their potential use for breeding of caffeine-free cultivars

Cryptic Diversity within Morphospecies\ud of Testate Amoebae (Amoebozoa:\ud Arcellinida) in New England Bogs\ud and Fens

Testate (shelled) amoebae are abundant and diverse in Sphagnum-rich areas of bogs and fens. Test morphology is standardly used to identify morphospecies as tests have varying shapes and compositions (e.g. siliceous, proteinaceous, agglutinated, or even calcareous). The recent application of molecular tools has revealed a greater complexity than morphology suggests, including multiple cryptic species. Here, we assess the biodiversity and relationships among eight morphospecies: Hyalosphenia elegans, Hyalosphenia papilio, Nebela carinata, Nebela flabellulum, Nebela militaris, Nebela tincta, Nebela tubulosa, and Quadrulella symmetrica using small subunit ribosomal DNA (SSU-rDNA). An SSU-rDNA phylogeny including 20 specimens from GenBank and 63 from this study reveals diversity within and among morphospecies and low resolution among some Nebela spp. Previous SSU-rDNA work on a limited sample of these species showed non-monophyly in the genus Hyalosphenia. Our analyses confirm this pattern and further suggest that other Nebela genera and morphospecies are not monophyletic. Moreover, inclusion of up to 24 specimens per species indicates non-monophyly of the morphospecies Hyalosphenia papilio and Hyalosphenia elegans. Our results suggest the morphological plasticity of testate amoebae across evolutionary time scales and that a combination of morphology and molecular analyses is needed to understand the biodiversity of these taxa.The authors would like to thank Jessica R. Grant, Jian Hua Lin, Adelaide Gordon, Charlene Gemora, Rose Katz-Berger for contributions to this research. Funding for the work comes from an NSF RUI Systematics grant (DEB RUI: 0919152). DJGL has been funded by CNPq GDE Fellowship 200853/2007-4 and a CNPq Post-Doctoral Fellowship 501089/2011-0. AMO was funded by Smith College Tomlinson Fund. Permits were obtained for all sites, from either National Park Service (Acadia) or Nature Conservancy (Hawley Bog