Regulation of cellular senescence by p53.

Many normal cells respond to potentially oncogenic stimuli by undergoing cellular senescence, a state of irreversibly arrested proliferation and altered differentiated function. Cellular senescence very likely evolved to suppress tumorigenesis. In support of this idea, it is regulated by several tumor suppressor genes. At the heart of this regulation is p53. p53 is essential for the senescence response to short telomeres, DNA damage, oncogenes and supraphysiological mitogenic signals, and overexpression of certain tumor suppressor genes. Despite the well-documented central role for p53 in the senescence response, many questions remain regarding how p53 senses senescence-inducing stimuli and how it elicits the senescent phenotype.     

Enhanced abundance of tintinnids under elevated CO<Subscript>2</Subscript> level from coastal Bay of Bengal

The role of microzooplankton (MZP) in the pelagic trophodynamics is highly significant, but the responses of marine MZP to increasing CO₂ levels are rather poorly understood. Hence the present study was undertaken to understand the responses of marine plankton to increasing CO₂ concentrations. Natural water samples from the coastal Bay of Bengal were incubated under the ambient condition and high CO₂ levels (703–711 μatm) for 5 days in May and June 2010. A significant negative correlation was obtained between phytoplankton and MZP abundance which indicated that phytoplankton community structure can considerably be controlled by MZP in this region. The average relative abundances of tintinnids under elevated CO₂ levels were found to be significantly higher (68.65 ± 5.63% in May; 85.46 ± 9.56% in June) than observed in the ambient condition (35.68 ± 6.83% in May; 79 ± 5.36% in June). The observed dominance of small chain forming diatom species probably played a crucial role as they can be potentially grazed by tintinnids. This fact was strengthened by the observed high negative correlations between the relative abundance of major phytoplankton and tintinnids. Moreover, particulate organic carbon and total bacterial counts were also enhanced under elevated CO₂ level and can serve as additional food source for ciliates. The observed responses of tintinnids to increasing CO₂ might have multiple impacts on the energy transfer, nutrient and carbon cycling in the coastal water. The duration of the present study was relatively short and therefore further investigation on longer time scale needs to be done and might give us a better insight about phytoplankton and MZP species succession under elevated CO₂ level.     

Ranging,Activity and Habitat Use by Tigers in the Mangrove Forests of the Sundarban

The Sundarban of India and Bangladesh (about 6000 km²) are the only mangrove forests inhabited by a sizeable population of tigers. The adjoining area also supports one of the highest human densities and experiences severe human-tiger conflicts. We used GPS-Satellite and VHF radio-collars on 6 (3 males and 3 female) tigers to study their ranging patterns and habitat preference. The average home range (95% Fixed Kernel) for resident females was 56.4 (SE 5.69) and for males it was 110 (SE 49) km². Tigers crossed an average of 5 water channels > 30 meters per day with a mean width of 54 meters, whereas channels larger than 400 meters were rarely crossed. Tigers spent over 58% of their time within Phoenix habitat but compositional analysis showed a habitat preference of the order Avicennia-Sonneratia > Phoenix > Ceriops > Barren > Water. Average daily distance moved was 4.6 km (range 0.1–23). Activity of tigers peaked between 05:00 hours and 10:00 hours showing some overlap with human activity. Territory boundaries were demarcated by large channels which tigers intensively patrolled. Extra caution should be taken while fishing or honey collection during early morning in Avicennia-Sonneratia and Phoenix habitat types along wide channels to reduce human-tiger conflict. Considering home-range core areas as exclusive, tiger density was estimated at 4.6 (SE range 3.6 to 6.7) tigers/100 km² giving a total population of 76 (SE range 59–110) tigers in the Indian Sundarban. Reluctance of tigers to cross wide water channels combined with increasing commercial boat traffic and sea level rise due to climate change pose a real threat of fragmenting the Sundarban tiger population.     

Physical mapping of repetitive extragenic palindromic sequences in Escherichia coli and phylogenetic distribution among Escherichia coli strains and other enteric bacteria.

Repetitive extragenic palindromic (REP) sequences are highly conserved inverted repeat sequences originally discovered in Escherichia coli and Salmonella typhimurium. We have physically mapped these sequences in the E. coli genome by using Southern hybridization of an ordered phage bank of E. coli (Y. Kohara, K. Akiyama, and K. Isono, Cell 50:495-508, 1987) with generic REP probes derived from the REP consensus sequence. The set of REP probe-hybridizing clones was correlated with a set of clones expected to contain REP sequences on the basis of computer searches. We also show that a generic REP probe can be used in Southern hybridization to analyze genomic DNA digested with restriction enzymes to determine genetic relatedness among natural isolates of E. coli. A search for these sequences in other members of the family Enterobacteriaceae shows a consistent correlation between both the number of occurrences and the hybridization strength and genealogical relationship.     

Distinct Roles for Rho Versus Rac/Cdc42 GTPases Downstream of Vav2 in Regulating Mammary Epithelial Acinar Architecture

Non-malignant mammary epithelial cells (MECs) undergo acinar morphogenesis in three-dimensional Matrigel culture, a trait that is lost upon oncogenic transformation. Rho GTPases are thought to play important roles in regulating epithelial cell-cell junctions, but their contributions to acinar morphogenesis remain unclear. Here we report that the activity of Rho GTPases is down-regulated in non-malignant MECs in three-dimensional culture with particular suppression of Rac1 and Cdc42. Inducible expression of a constitutively active form of Vav2, a Rho GTPase guanine nucleotide exchange factor activated by receptor tyrosine kinases, in three-dimensional MEC culture activated Rac1 and Cdc42; Vav2 induction from early stages of culture impaired acinar morphogenesis, and induction in preformed acini disrupted the pre-established acinar architecture and led to cellular outgrowths. Knockdown studies demonstrated that Rac1 and Cdc42 mediate the constitutively active Vav2 phenotype, whereas in contrast, RhoA knockdown intensified the Vav2-induced disruption of acini, leading to more aggressive cell outgrowth and branching morphogenesis. These results indicate that RhoA plays an antagonistic role to Rac1/Cdc42 in the control of mammary epithelial acinar morphogenesis.     

Isolation and characterization of a novel DNA methyltransferase complex linking DNMT3B with components of the mitotic chromosome condensation machinery

Proper patterns of genome-wide DNA methylation, mediated by DNA methyltransferases DNMT1, -3A and -3B, are essential for embryonic development and genomic stability in mammalian cells. The de novo DNA methyltransferase DNMT3B is of particular interest because it is frequently overexpressed in tumor cells and is mutated in immunodeficiency, centromere instability and facial anomalies (ICF) syndrome. In order to gain a better understanding of DNMT3B, in terms of the targeting of its methylation activity and its role in genome stability, we biochemically purified endogenous DNMT3B from HeLa cells. DNMT3B co-purifies and interacts, both in vivo and in vitro, with several components of the condensin complex (hCAP-C, hCAP-E and hCAP-G) and KIF4A. Condensin mediates genome-wide chromosome condensation at the onset of mitosis and is critical for proper segregation of sister chromatids. KIF4A is proposed to be a motor protein carrying DNA as cargo. DNMT3B also interacts with histone deacetylase 1 (HDAC1), the co-repressor SIN3A and the ATP-dependent chromatin remodeling enzyme hSNF2H. Further more, DNMT3B co-localizes with condensin and KIF4A on condensed chromosomes throughout mitosis. These studies therefore reveal the first direct link between the machineries regulating DNA methylation and mitotic chromosome condensation in mammalian cells.     

Cbl-mediated ubiquitinylation is required for lysosomal sorting of epidermal growth factor receptor but is dispensable for endocytosis

Ligand-induced down-regulation controls the signaling potency of the epidermal growth factor receptor (EGFR/ErbB1). Overexpression studies have identified Cbl-mediated ubiquitinylation of EGFR as a mechanism of ligand-induced EGFR down-regulation. However, the role of endogenous Cbl in EGFR down-regulation and the precise step in the endocytic pathway regulated by Cbl remain unclear. Using Cbl-/- mouse embryonic fibroblast cell lines, we demonstrate that endogenous Cbl is essential for ligand-induced ubiquitinylation and efficient degradation of EGFR. Further analyses using Chinese hamster ovary cells with a temperature-sensitive defect in ubiquitinylation confirm a crucial role of the ubiquitin machinery in Cbl-mediated EGFR degradation. However, internalization into early endosomes did not require Cbl function or an intact ubiquitin pathway. Confocal immunolocalization studies indicated that Cbl-dependent ubiquitinylation plays a critical role at the early endosome to late endosome/lysosome sorting step of EGFR down-regulation. These findings establish Cbl as the major endogenous ubiquitin ligase responsible for EGFR degradation, and show that the critical role of Cbl-mediated ubiquitinylation is at the level of endosomal sorting, rather than at the level of internalization.