First description of nest-decoration behaviour in a wild sub-Antarctic shorebird.

A wide range of animal species accumulate objects in, on, and/or around structures they build. Sometimes, these accumulations serve specific functions (e.g. structural or isolating features) or are purely incidental, while in other cases the materials are deliberately displayed to serve signalling purposes (extended phenotype signals). In this pilot study, we employed systematic in situ observations and camera trapping to describe for the first time that both partners of a territorial shorebird, the black-faced sheathbill (Chionis minor ssp minor) collect, carry, and arrange colourful marine shells and dry twigs within and around their nest cavity. Our observations expand the taxonomic breadth of avian extended phenotype signals, by showing that at least one species within a largely understudied group i.e., Charadriiformes, exhibits nest-decoration behaviour. Multiple manipulative experiments are needed to explore further the signalling function of these decorations, which opens new exciting avenues for animal communication and cognition research.

Nest design in a changing world: great tit Parus major nests from a Mediterranean city environment as a case study.

Investigations of urbanization effects on birds have focused mainly on breeding traits expressed after the nest-building stage (e.g. first-egg date, clutch size, breeding success, and offspring characteristics). Urban studies largely ignored how and why the aspects of nest building might be associated with the degree of urbanization. As urban environments are expected to present novel environmental changes relative to rural environments, it is important to evaluate how nest-building behavior is impacted by vegetation modifications associated with urbanization. To examine nest design in a Mediterranean city environment, we allowed urban great tits (Parus major) to breed in nest boxes in areas that differed in local vegetation cover. We found that different measures of nest size or mass were not associated with vegetation cover. In particular, nests located adjacent to streets with lower vegetation cover were not smaller or lighter than nests in parks with higher vegetation cover. Nests adjacent to streets contained more pine needles than nests in parks. In addition, in nests adjacent to streets, nests from boxes attached to pine trees contained more pine needles than nests from boxes attached to other trees. We suggest that urban-related alterations in vegetation cover do not directly impose physical limits on nest size in species that are opportunistic in the selection of nesting material. However, nest composition as reflected in the use of pine needles was clearly affected by habitat type and the planted tree species present, which implies that rapid habitat change impacts nest composition. We do not exclude that urbanization might impact other aspects of nest building behaviour not covered in our study (e.g. costs of searching for nest material), and that the strengths of the associations between urbanization and nest structures might differ among study populations or species.

Transcriptional repression of estrogen receptor α signaling by SENP2 in breast cancer cells.

Estrogen receptors (ERs) are ligand-activated transcription factors involved in many physiological and pathological processes, including breast cancer. Their activity is fine-tuned by posttranslational modifications, notably sumoylation. In the present study, we investigated the role of the small ubiquitin-related modifier (SUMO) protease, SUMO1/sentrin/suppressor of Mif 2-specific peptidase 2 (SENP2), in the regulation of ERα activity. We first found SENP2 to significantly repress estradiol-induced transcriptional activity in breast cancer cells (MCF7 and T47D). This effect was observed with a reporter plasmid and on endogenous genes such as TFF1 and CTSD, which were shown to recruit SENP2 in chromatin immunoprecipitation experiments. Using glutathione S-transferase pull-down, coimmunoprecipitation and proximity ligation assays, SENP2 was found to interact with ERα and this interaction to be mediated by the amino-terminal region of the protease and the hinge region of the receptor. Interestingly, we demonstrated that ERα repression by SENP2 is independent of its SUMO protease activity and requires a transcriptional repressive domain located in the amino-terminal end of the protease. Using small interfering RNA assays, we evidenced that this domain recruits the histone deacetylase 3 (HDAC3), to be fully active. Furthermore, using both overexpression and knockdown strategies, we showed that SENP2 robustly represses estrogen-dependent and independent proliferation of MCF7 cells. We provided evidence that this effect requires both the proteolytic and transcriptional activities of SENP2. Altogether, our study unravels a new property for a SUMO protease and identifies SENP2 as a classical transcription coregulator.

Receptor-interacting protein 140 is a repressor of the androgen receptor activity.

The androgen receptor (AR) is a ligand-activated transcription factor that controls growth and survival of prostate cancer cells. In the present study, we investigated the regulation of AR activity by the receptor-interacting protein 140 (RIP140). We first showed that RIP140 could be coimmunoprecipitated with the receptor when coexpressed in 293T cells. This interaction appeared physiologically relevant because chromatin immunoprecipitation assays revealed that, under R1881 treatment, RIP140 could be recruited to the prostate-specific antigen encoding gene in LNCaP cells. In vitro glutathione S-transferase pull-down assays provided evidence that the carboxy-terminal domain of AR could interact with different regions of RIP140. By means of fluorescent proteins, we demonstrated that ligand-activated AR was not only able to translocate to the nucleus but also to relocate RIP140 from very structured nuclear foci to a diffuse pattern. Overexpression of RIP140 strongly repressed AR-dependent transactivation by preferentially targeting the ligand binding domain-dependent activity. Moreover, disruption of RIP140 expression induced AR overactivation, thus revealing RIP140 as a strong AR repressor. We analyzed its mechanism of transrepression and first demonstrated that different regions of RIP140 could mediate AR-dependent repression. We then showed that the carboxy-terminal end of RIP140 could reverse transcriptional intermediary factor 2-dependent overactivation of AR. The use of mutants of RIP140 allowed us to suggest that C-terminal binding protein played no role in RIP140-dependent inhibition of AR activity, whereas histone deacetylases partly regulated that transrepression. Finally, we provided evidence for a stimulation of RIP140 mRNA expression in LNCaP cells under androgen treatment, further emphasizing the role of RIP140 in androgen signaling.

Identification of genes involved in growth inhibition of breast cancer cells transduced with estrogen receptor.

Estrogen receptor alpha (ERalpha)-negative breast cancer cells display an aggressive phenotype. We previously showed that adenoviral expression of ERalpha in ER-negative breast cancer cells leads to an estrogen-dependent down-regulation of the proliferation, which could be of interest to control the growth of such cells. In this study, we observed an increase in protein levels of p21 and p27 cyclin-dependent kinase inhibitors, whereas pRb phosphorylation is strongly decreased. Flow cytometry experiments showed a slower transit of cells in G1 (hormone-independent), a hormone-induced accelerated transit through S phase and a possible arrest in G2/M phase. In addition, ERalpha-expressing cells were undergoing apoptosis. By using cDNA macroarrays, we identified a novel collection of genes regulated by liganded ERalpha potentially regulating cell cycle, apoptosis, cell signalling, stress response and DNA repair.

Comparative transductions of breast cancer cells by three DNA viruses.

Defining the ideal vectors to transduce breast cancer using viruses is currently under intense pre-clinical evaluation. Our study constitutes the first direct comparison of the infection efficiencies of a human serotype 5 (Ad5), a canine serotype 2 (CAV-2) adenovirus, and a human serotype 2 adeno-associated virus (AAV-2) in breast cancer cells. We observed an excellent infection efficiency for Ad5 vector, whereas both CAV-2 and AAV-2 vectors lead to low infection of these cells. Real-time PCR, flow cytometry, and antibody blocking studies suggest that Ad5 and CAV-2 infection ability is not strictly dependent on coxsackie adenovirus receptor (CAR) or alpha(v) integrin levels. In conclusion, our data suggest that human adenoviruses are excellent transducers of breast cancer cells, though it may be difficult to predict the extent of infection solely on CAR or alpha(v) integrin levels.

IL-8 expression and its possible relationship with estrogen-receptor-negative status of breast cancer cells.

Estrogen-receptor (ER) status is an important parameter in breast cancer management as ER-positive breast cancers have a better prognosis than ER-negative tumors. This difference comes essentially from the lower aggressiveness and invasiveness of ER-positive tumors. Here, we demonstrate, that interleukin-8 (IL-8) was clearly overexpressed in most ER-negative breast, ovary cell lines and breast tumor samples tested, whereas no significant IL-8 level could be detected in ER-positive breast or ovarian cell lines. We have also cloned human IL-8 from ER-negative MDA-MB-231 cells, and we show that IL-8 produced by breast cancer cells is identical to monocyte-derived IL-8. Interestingly, the invasion potential of ER-negative breast cancer cells is associated at least in part with expression of IL-8, but not with IL-8 receptor levels. Moreover, IL-8 increases the invasiveness of ER-positive breast cancer cells by two fold, thus confirming the invasion-promoting role of IL-8. On the other hand, exogenous expression of estrogen receptors in ER-negative cells led to a decrease of IL-8 levels. In summary, our data show that IL-8 expression is negatively linked to ER status of breast and ovarian cancer cells. We also support the idea that IL-8 expression is associated with a higher invasiveness potential of cancer cells in vitro, which suggests that IL-8 could be a novel marker of tumor aggressiveness.

Procathepsin D interacts with prosaposin in cancer cells but its internalization is not mediated by LDL receptor-related protein.

The cell surface binding, endocytosis, and lysosomal routing of procathepsin D (procath-D) in cancer cells are mostly independent of the mannose-6-phosphate (M6P) receptors. In an attempt to define the receptor involved, we intracellularly cross-linked procath-D with a 68-kDa protein that we identified with specific antibodies as prosaposin in human breast and ovarian cancer cell lines. In cancer cells, this protein-protein interaction was resistant to ammonium chloride or M6P treatment, indicating that it was independent of the M6P receptors. A similar interaction also occurred in the breast cancer cell culture medium between the secreted prosaposin and procath-D. Since these two precursors can be endocytosed, we then determined whether they were interacting with the same cell surface receptor. In fibroblasts, we confirmed that the endocytosis of these two proteins was different since it was generally mediated by the M6P receptors for procath-D and mostly by LRP (LDL receptor-related protein) for prosaposin. In breast cancer cells, prosaposin endocytosis was not detected, in contrast to procath-D endocytosis, suggesting that the majority of procath-D is not internalized as a complex with prosaposin. Moreover, RAP (receptor-associated protein), a ligand inhibiting LRP-mediated endocytosis, prevented internalization of prosaposin in 49-F rat fibroblasts, but did not affect procath-D M6P-independent internalization in MDA-MB231 cells. We conclude that in breast cancer cells, even though procath-D interacts intracellularly and extracellarly with prosaposin, it is endocytosed independent of prosaposin by a receptor different from the M6P receptors and the LRP.