Yeast Screens Identify the RNA Polymerase II CTD and SPT5 as Relevant Targets of BRCA1 Interaction

BRCA1 has been implicated in numerous DNA repair pathways that maintain genome integrity, however the function responsible for its tumor suppressor activity in breast cancer remains obscure. To identify the most highly conserved of the many BRCA1 functions, we screened the evolutionarily distant eukaryote Saccharomyces cerevisiae for mutants that suppressed the G1 checkpoint arrest and lethality induced following heterologous BRCA1 expression. A genome-wide screen in the diploid deletion collection combined with a screen of ionizing radiation sensitive gene deletions identified mutants that permit growth in the presence of BRCA1. These genes delineate a metabolic mRNA pathway that temporally links transcription elongation (SPT4, SPT5, CTK1, DEF1) to nucleopore-mediated mRNA export (ASM4, MLP1, MLP2, NUP2, NUP53, NUP120, NUP133, NUP170, NUP188, POM34) and cytoplasmic mRNA decay at P-bodies (CCR4, DHH1). Strikingly, BRCA1 interacted with the phosphorylated RNA polymerase II (RNAPII) carboxy terminal domain (P-CTD), phosphorylated in the pattern specified by the CTDK-I kinase, to induce DEF1-dependent cleavage and accumulation of a RNAPII fragment containing the P-CTD. Significantly, breast cancer associated BRCT domain defects in BRCA1 that suppressed P-CTD cleavage and lethality in yeast also suppressed the physical interaction of BRCA1 with human SPT5 in breast epithelial cells, thus confirming SPT5 as a relevant target of BRCA1 interaction. Furthermore, enhanced P-CTD cleavage was observed in both yeast and human breast cells following UV-irradiation indicating a conserved eukaryotic damage response. Moreover, P-CTD cleavage in breast epithelial cells was BRCA1-dependent since damage-induced P-CTD cleavage was only observed in the mutant BRCA1 cell line HCC1937 following ectopic expression of wild type BRCA1. Finally, BRCA1, SPT5 and hyperphosphorylated RPB1 form a complex that was rapidly degraded following MMS treatment in wild type but not BRCA1 mutant breast cells. These results extend the mechanistic links between BRCA1 and transcriptional consequences in response to DNA damage and suggest an important role for RNAPII P-CTD cleavage in BRCA1-mediated cancer suppression

Why Functional Pre-Erythrocytic and Bloodstage Malaria Vaccines Fail: A Meta-Analysis of Fully Protective Immunizations and Novel Immunological Model

Background: Clinically protective malaria vaccines consistently fail to protect adults and children in endemic settings, and at best only partially protect infants. Methodology/Principal Findings: We identify and evaluate 1916 immunization studies between 1965-February 2010, and exclude partially or nonprotective results to find 177 completely protective immunization experiments. Detailed reexamination reveals an unexpectedly mundane basis for selective vaccine failure: live malaria parasites in the skin inhibit vaccine function. We next show published molecular and cellular data support a testable, novel model where parasite-host interactions in the skin induce malaria-specific regulatory T cells, and subvert early antigen-specific immunity to parasite-specific immunotolerance. This ensures infection and tolerance to reinfection. Exposure to Plasmodium-infected mosquito bites therefore systematically triggers immunosuppression of endemic vaccine-elicited responses. The extensive vaccine trial data solidly substantiate this model experimentally. Conclusions/Significance: We conclude skinstage-initiated immunosuppression, unassociated with bloodstage parasites, systematically blocks vaccine function in the field. Our model exposes novel molecular and procedural strategies to significantly and quickly increase protective efficacy in both pipeline and currently ineffective malaria vaccines, and forces fundamental reassessment of central precepts determining vaccine development. This has major implications fo

The role of cGMP signalling in regulating life cycle progression of Plasmodium.

The 3'-5'-cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG) is the main mediator of cGMP signalling in the malaria parasite. This article reviews the role of PKG in Plasmodium falciparum during gametogenesis and blood stage schizont rupture, as well as the role of the Plasmodium berghei orthologue in ookinete differentiation and motility, and liver stage schizont development. The current views on potential effector proteins downstream of PKG and the mechanisms that may regulate cyclic nucleotide levels are presented

Petrogenetic and Compositional Features of Rare Metal PanAfrican Post-Collisional Pegmatites of Southwestern Nigeria; A Status Review

This research reviews the geology, petrogenesis, compositional trends and geochronology of the rare-metal pegmatite of southwestern Nigeria. The source of these pegmatites is still presently debated which have been explained as either product of highly fractionated molten material or anatexis of the local crust. However, published works of past authors have been compiled to give a detailed understanding of the formation of the mineral deposits. The basement complex of southwestern Nigeria comprises of Precambrian rocks of amphibolite, the hornblende gneiss and the granite gneisses which were formed as a result of the opening and closing of the ensialic basin with significant, extensive subduction during the Pan-African orogeny. The pegmatites in this region have shown internal zoning and a high degree of evolution from the border zone to the core zone during the crystallization and solidification of the felsic granite to pegmatite melt. The rare-metal pegmatites have distinct chemical compositions and mineralogy, containing quartz, biotite, muscovite, microcline, garnet with localized tourmaline, tantalite and columbite. These pegmatites vary significantly by their bulk-rock and mineral chemistry which indicates a more peraluminous attribute and enrichments of lithophile elements of Rb, Cs, Ta and Ba. Previous K/Ar isotopic ages (502.8±13.0 Ma and 514.5±13.2 Ma) suggest that the pegmatites are related to the post-collisional phase of intensive metasomatism. Adopted from previous studies, a five-stage conceptual model of evolution which is widely accepted have been proposed for the origin of the pegmatites

Geoelectrical Characterization of Coastal Aquifers in Agbado-Ijaye, Lagos, Southwestern Nigeria; Implications for Groundwater Resources Sustainability

Water is a natural resource; its availability depends on climatic and geological conditions, and it is invariably controlled by human activities. Agbado-Ijaye lies within a coastal area, where local communities have been facing incessant water shortages, especially during the dry season. This study investigated the groundwater-bearing geological unit(s) using hydrogeophysical techniques in the coastal environment. The electrical resistivity technique, involving vertical electrical sounding (VES) and two-dimensional (2D) electrical resistivity imaging via Wenner array electrode configuration, was used to characterize the geoelectric distribution. Twenty VES stations were investigated and current electrodes (AB/2 m) spacing expanded from 1–200 m; four 2D electrical resistivity imaging traverses having a length of 200 m each and interelectrode spacing of 10 m (level 1) to 60 m (level 6) was adopted. Four geoelectric units were delineated, namely: topsoil (15–251 Ωm), clayey (28–100 Ωm), clayey sand (125–190 Ωm) and sandy (205–876 Ωm) with thicknesses ranging from 0.7–1.3 m, 4.1–19.0 m, 2.6–15.6 m and undefined depth, respectively. The 2D imaging sections also detected similar geoelectric layers, corroborating the VES-derived sections. The inverted sections delineated two different aquifers: the shallower low-yield aquifer comprising sandy clay/clayey sand units with a maximum depth of about 5.5 m. This layer is adjudged to be the continental plain sand of the Benin Formation. The deeper high-yield aquifer with a maximum depth of 30.4 m is a beach sand unit that belongs to the Tertiary Alluvium of the Dahomey Basin. The study showed that hydrogeophysical investigation is vital in exploring, developing, and managing coastal groundwater resources.</jats:p

Geoelectrical Characterization of Coastal Aquifers in Agbado-Ijaye, Lagos, Southwestern Nigeria; Implications for Groundwater Resources Sustainability

Water is a natural resource; its availability depends on climatic and geological conditions, and it is invariably controlled by human activities. Agbado-Ijaye lies within a coastal area, where local communities have been facing incessant water shortages, especially during the dry season. This study investigated the groundwater-bearing geological unit(s) using hydrogeophysical techniques in the coastal environment. The electrical resistivity technique, involving vertical electrical sounding (VES) and two-dimensional (2D) electrical resistivity imaging via Wenner array electrode configuration, was used to characterize the geoelectric distribution. Twenty VES stations were investigated and current electrodes (AB/2 m) spacing expanded from 1–200 m; four 2D electrical resistivity imaging traverses having a length of 200 m each and interelectrode spacing of 10 m (level 1) to 60 m (level 6) was adopted. Four geoelectric units were delineated, namely: topsoil (15–251 Ωm), clayey (28–100 Ωm), clayey sand (125–190 Ωm) and sandy (205–876 Ωm) with thicknesses ranging from 0.7–1.3 m, 4.1–19.0 m, 2.6–15.6 m and undefined depth, respectively. The 2D imaging sections also detected similar geoelectric layers, corroborating the VES-derived sections. The inverted sections delineated two different aquifers: the shallower low-yield aquifer comprising sandy clay/clayey sand units with a maximum depth of about 5.5 m. This layer is adjudged to be the continental plain sand of the Benin Formation. The deeper high-yield aquifer with a maximum depth of 30.4 m is a beach sand unit that belongs to the Tertiary Alluvium of the Dahomey Basin. The study showed that hydrogeophysical investigation is vital in exploring, developing, and managing coastal groundwater resources