Identification and Characterization of Nucleolin as a COUP-TFII Coactivator of Retinoic Acid Receptor β Transcription in Breast Cancer Cells

The orphan nuclear receptor COUP-TFII plays an undefined role in breast cancer. Previously we reported lower COUP-TFII expression in tamoxifen/endocrine-resistant versus sensitive breast cancer cell lines. The identification of COUP-TFII-interacting proteins will help to elucidate its mechanism of action as a transcriptional regulator in breast cancer.FLAG-affinity purification and multidimensional protein identification technology (MudPIT) identified nucleolin among the proteins interacting with COUP-TFII in MCF-7 tamoxifen-sensitive breast cancer cells. Interaction of COUP-TFII and nucleolin was confirmed by coimmunoprecipitation of endogenous proteins in MCF-7 and T47D breast cancer cells. In vitro studies revealed that COUP-TFII interacts with the C-terminal arginine-glycine repeat (RGG) domain of nucleolin. Functional interaction between COUP-TFII and nucleolin was indicated by studies showing that siRNA knockdown of nucleolin and an oligonucleotide aptamer that targets nucleolin, AS1411, inhibited endogenous COUP-TFII-stimulated RARB2 expression in MCF-7 and T47D cells. Chromatin immunoprecipitation revealed COUP-TFII occupancy of the RARB2 promoter was increased by all-trans retinoic acid (atRA). RARβ2 regulated gene RRIG1 was increased by atRA and COUP-TFII transfection and inhibited by siCOUP-TFII. Immunohistochemical staining of breast tumor microarrays showed nuclear COUP-TFII and nucleolin staining was correlated in invasive ductal carcinomas. COUP-TFII staining correlated with ERα, SRC-1, AIB1, Pea3, MMP2, and phospho-Src and was reduced with increased tumor grade.Our data indicate that nucleolin plays a coregulatory role in transcriptional regulation of the tumor suppressor RARB2 by COUP-TFII

Reproductive biology of Echinometra lucunter (Echinodermata: Echinoidea) in a northeast Brazilian sandstone reef

The edible sea urchin Echinometra lucunter (Linnaeus, 1758) is a very common species on the sublittoral-midlittoral in Brazilian rocky shores. The aim of this work was to describe the gametogenesis and reproductive strategy of the E. lucunter population at Muro Alto beach in the Northeast coast of Brazil from August 2004 to August 2005. A total of 240 specimens were collected on the sandstone reef flat from a tidepool during spring low tides. The overall sex ratio was1.12:1,withoutsignificanttemporalvariationexceptinOctober2004. Firsts sexual maturity ocurred in individuals from a diameter of 20.8 mm. There was not a significant difference in gonad index between females and males during the sampling period. The female's gonad index variation was associated with a well-defined spawning, corroborated by the histological analysis of the gonads, which demonstrates sex differences of the gamete production. By contrast, the males showed no clear pattern. It is suggested that continuous reproduction with seasonal peaks in the E. lucunter population occurs at Muro Alto beach.<br>O ouriço-do-mar comestível Echinometra lucunter (Linnaeus, 1758) é uma espécie muito comum no infralitoral e mediolitoral do Brasil. O objetivo deste trabalho foi descrever a gametogênese e a estratégia reprodutiva da população de E. lucunterna praia de Muro Alto entre agosto de 2004 e agosto de 2005. Um total de 240 espécimes foi capturado de uma poça de maré situada no topo recifal, durante as marés baixas de sizígia. A razão sexual total foi de 1,12:1 sem variação temporal significativa, exceto em outubro de 2004. A primeira maturidade sexual ocorreu em indivíduos a partir de 20,8 mm de diâmetro. Não houve nenhuma diferença significativa no índice gonadal entre fêmeas e machos durante o período de amostragem. A variação do índice gonadal das fêmeas foi associada a um período de desova bem definido, corroborado pela análise histológica das gônadas, que demonstra diferenças sexuais na produção de gametas. Ao contrário, os machos não apresentaram nenhum padrão. Sugere-se que a reprodução da população de E. lucunter na praia de Muro Alto é contínua, compicos sazonais

Estimating rates of biologically driven coral reef framework production and erosion: a new census-based carbonate budget methodology and applications to the reefs of Bonaire

Census-based approaches can provide important measures of the ecological processes controlling reef carbonate production states. Here, we describe a rapid, non-destructive approach to carbonate budget assessments, termed ReefBudget that is census-based and which focuses on quantifying the relative contributions made by different biological carbonate producer/eroder groups to net reef framework carbonate production. The methodology is presently designed only for Caribbean sites, but has potential to be adapted for use in other regions. Rates are calculated using data on organism cover and abundance, combined with annual extension or production rate measures. Set against this are estimates of the rates at which bioeroding species of fish, urchins and internal substrate borers erode reef framework. Resultant data provide a measure of net rates of biologically driven carbonate production (kg CaCO3 m(-2) year(-1)). These data have potential to be integrated into ecological assessments of reef state, to aid monitoring of temporal (same-site) changes in rates of biological carbonate production and to provide insights into the key ecological drivers of reef growth or erosion as a function of environmental change. Individual aspects of the budget methodology can also be used alongside other census approaches if deemed appropriate for specific study aims. Furthermore, the methodology spreadsheets are user-changeable, allowing local or new process/rate data to be integrated into calculations. Application of the methodology is considered at sites around Bonaire. Highest net rates of carbonate production, +9.52 to +2.30 kg CaCO3 m(-2) year(-1), were calculated at leeward sites, whilst lower rates, +0.98 to -0.98 kg CaCO3 m(-2) year(-1), were calculated at windward sites. Data are within the ranges calculated in previous budget studies and provide confidence in the production estimates the methodology generates