Subnanometre-resolution structure of the intact Thermus thermophilus H+-driven ATP synthase

Ion-translocating rotary ATPases serve either as ATP synthases, using energy from a transmembrane ion motive force to create the cell's supply of ATP, or as transmembrane ion pumps that are powered by ATP hydrolysis. The members of this family of enzymes each contain two rotary motors: one that couples ion translocation to rotation and one that couples rotation to ATP synthesis or hydrolysis. During ATP synthesis, ion translocation through the membrane-bound region of the complex causes rotation of a central rotor that drives conformational changes and ATP synthesis in the catalytic region of the complex. There are no structural models available for the intact membrane region of any ion-translocating rotary ATPase. Here we present a 9.7?? resolution map of the H(+)-driven ATP synthase from Thermus thermophilus obtained by electron cryomicroscopy of single particles in ice. The 600-kilodalton complex has an overall subunit composition of A(3)B(3)CDE(2)FG(2)IL(12). The membrane-bound motor consists of a ring of L subunits and the carboxy-terminal region of subunit I, which are equivalent to the c and a subunits of most other rotary ATPases, respectively. The map shows that the ring contains 12 L subunits and that the I subunit has eight transmembrane helices. The L(12) ring and I subunit have a surprisingly small contact area in the middle of the membrane, with helices from the I subunit making contacts with two different L subunits. The transmembrane helices of subunit I form bundles that could serve as half-channels across the membrane, with the first half-channel conducting protons from the periplasm to the L(12) ring and the second half-channel conducting protons from the L(12) ring to the cytoplasm. This structure therefore suggests the mechanism by which a transmembrane proton motive force is converted to rotation in rotary ATPases.     

Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling

The adult stem cell marker Lgr5 and its relative Lgr4 are often co-expressed in Wnt-driven proliferative compartments. We find that conditional deletion of both genes in the mouse gut impairs Wnt target gene expression and results in the rapid demise of intestinal crypts, thus phenocopying Wnt pathway inhibition. Mass spectrometry demonstrates that Lgr4 and Lgr5 associate with the Frizzled/Lrp Wnt receptor complex. Each of the four R-spondins, secreted Wnt pathway agonists, can bind to Lgr4, -5 and -6. In HEK293 cells, RSPO1 enhances canonical WNT signals initiated by WNT3A. Removal of LGR4 does not affect WNT3A signalling, but abrogates the RSPO1-mediated signal enhancement, a phenomenon rescued by re-expression of LGR4, -5 or -6. Genetic deletion of Lgr4/5 in mouse intestinal crypt cultures phenocopies withdrawal of Rspo1 and can be rescued by Wnt pathway activation. Lgr5 homologues are facultative Wnt receptor components that mediate Wnt signal enhancement by soluble R-spondin proteins. These results will guide future studies towards the application of R-spondins for regenerative purposes of tissues expressing Lgr5 homologues.     

Systematic identification of genomic markers of drug sensitivity in cancer cells

Clinical responses to anticancer therapies are often restricted to a subset of patients. In some cases, mutated cancer genes are potent biomarkers for responses to targeted agents. Here, to uncover new biomarkers of sensitivity and resistance to cancer therapeutics, we screened a panel of several hundred cancer cell lines--which represent much of the tissue-type and genetic diversity of human cancers--with 130 drugs under clinical and preclinical investigation. In aggregate, we found that mutated cancer genes were associated with cellular response to most currently available cancer drugs. Classic oncogene addiction paradigms were modified by additional tissue-specific or expression biomarkers, and some frequently mutated genes were associated with sensitivity to a broad range of therapeutic agents. Unexpected relationships were revealed, including the marked sensitivity of Ewing's sarcoma cells harbouring the EWS (also known as EWSR1)-FLI1 gene translocation to poly(ADP-ribose) polymerase (PARP) inhibitors. By linking drug activity to the functional complexity of cancer genomes, systematic pharmacogenomic profiling in cancer cell lines provides a powerful biomarker discovery platform to guide rational cancer therapeutic strategies.     

Quality vs. quantity of publications in nanotechnology field from the People’s Republic of China

This study evaluates trends in quality of nanotechnology and nanoscience papers produced by authors from the People＇s Republic of China （PRC）. The metric used to gauge quality is ratio of highly cited nanotechnology papers to total nanotechnology papers produced in sequential time frames. The USA is both the most prolific nanotechnology publishing country and most represented country on highly cited nanotechnology papers （both in absolute numbers of highly cited papers and highly cited papers relative to total publications） over the 1998-2003 time frame, based on the SCl/SSCl databases. Some of the smaller hi-tech countries have relatively high ratios （-2） of highly cited papers to total publications （e.g. Denmark, Netherlands, Switzerland）. Countries that have exhibited rapid growth in SCl/SSCl nanotechnology paper production in recent years （e.g. PRC, South Korea） had ratios an order of magnitude less than that of the USA for 1998, but by 2003 had increased to about 20% that of the USA （-2.5）. PRC and South Korea have climbed in the publications rankings from 6th and 9th in 1998, respectively, to 2nd and 6th in 2005, respectively. PRC＇s ratio monotonically increased from 0.16 to 0.45 over the 1998-2003 period, and South Korea＇s ratio increased from 0.11 to about 0.6 over that same period, indicating their papers are getting more and more citations proportionately. Thus, under rapid growth conditions, PRC and South Korea have been able to increase their share of participation in highly cited papers. As of 2003, PRC and South Korea have ratios comparable to nations like Japan, France, Italy, and Australia but not yet approaching those of the highly cited countries. None of the top ten publications producing institutions are from the USA, while all of the top ten highly cited publications producers are from the USA. Over the 1998-2003 time period, the top six total publications producing institutions （globally） remained the same, with Chinese Academy of Sciences （which consists