Androgen regulation of cell proliferation and expression of viral sequences in mouse mammary tumour cells

The role of steroids in promoting cell proliferation is well established but the molecular mechanisms are not clear. The S115 mouse mammary tumour cell line provides a model system for molecular studies in vitro in that it exhibits in tissue culture both a positive proliferative response to androgens and a change from a transformed phenotype in the presence of androgen to a normal phenotype when androgen is removed. We have considered here the possible involvement of mouse mammary tumour virus (MMTV) in these processes. We have demonstrated the presence in S115 cells of MMTV-related sequences which are transcribed into RNA only in the long-term presence of androgen. Prolonged culture in the absence of androgen, which results in loss of proliferative response to androgen, is accompanied by loss of MMTV-related RNA and increased methylation of MMTV-related sequences.     

Conformational flexibility of poly (dG-m5dC) under very low salt conditions

Summary The methylated DNA polymer poly (dG-m⁵dC) which exhibits a B helical conformation in solutions containing 20 mM NaCl, undergoes a gradual and reversible transition to the Z conformation as the NaCl concentration is lowered. The midpoint of this transition occurs around 5–6 mM NaCl. The conformational flexibility of this polymer at such low NaCl concentrations opens up the possibility of studying the effects of other perturbants with negligible interference from salt concentration effects.     

Reactivity of HTLV-transformed human T-cell lines to MHC class II antigens

T-cell lines established from individuals infected with human T-cell leukaemia virus (HTLV) or generated by co-cultivation of normal human T cells with HTLV-infected T-cells, express class II (HLA-D/DR or Ia) antigens of the major histocompatibility complex (MHC) and interleukin-2 (IL-2) receptors. Because the expression of these markers characterizes the differentiation of immunologically activated T cells, we have now explored the possibility that HTLV- infected T cells might be primed to autologous or allogeneic Ia antigens expressed by the infecting cells. Our studies on the capacity of HTLV-infected T cells to display responses on mixed lymphocyte culture indicate that such T cells as well as single-cell clones derived from them, react non-discriminatively to all known allelic variants of human HLA-D/DR antigens, including those expressed by the responding cells. This reaction is inhibited by antibody to human Ia and is not triggered by Ia-negative T-leukaemia cells. The structure recognized seems to be a common epitope determinant of human Ia antigens, as (HTLV-infected) T cells primed in vitro to one HLA-D/DR specificity display amplified responses to all other HLA-D/DR antigens. We therefore believe that autostimulation by a self-Ia determinant may trigger the clonal expansion of HTLV-infected T cells and potentiate autoimmune processes.     

The effect of alkylation on the antimicrobial activities of 7-hydroxy- and 4-hydroxycoumarins

Zusammenfassung Es wird der Einfluss einer Alkylierung auf die antimikrobielle Wirkung von 7-Hydroxy- und 4-Hydroxy-Cumarin dargestellt.     

Electron-microscopic mapping of the hinge region of myosin

Summary The trypsin-sensitive sites in the labile hinge region of the myosin molecule are located with heightened accuracy (±2 nm) by electron microscopy as lying at 70, 85, 95, and 103 nm from the C-terminus of the rod section of the molecule.I thank Dr.Michael Young for valuable advice and encouragement, Drs.Jerome Gross andRomaine R. Bruns for use of their electron microscope, and Mrs.Muriel H. Blanchard for excellent technical assistance. This work was supported by the American Heart Association, The John A. Hartford Foundation, Inc., and the National Institutes of Health.     

Backbone structure of the infectious epsilon15 virus capsid revealed by electron cryomicroscopy

A half-century after the determination of the first three-dimensional crystal structure of a protein, more than 40,000 structures ranging from single polypeptides to large assemblies have been reported. The challenge for crystallographers, however, remains the growing of a diffracting crystal. Here we report the 4.5-A resolution structure of a 22-MDa macromolecular assembly, the capsid of the infectious epsilon15 (epsilon15) particle, by single-particle electron cryomicroscopy. From this density map we constructed a complete backbone trace of its major capsid protein, gene product 7 (gp7). The structure reveals a similar protein architecture to that of other tailed double-stranded DNA viruses, even in the absence of detectable sequence similarity. However, the connectivity of the secondary structure elements (topology) in gp7 is unique. Protruding densities are observed around the two-fold axes that cannot be accounted for by gp7. A subsequent proteomic analysis of the whole virus identifies these densities as gp10, a 12-kDa protein. Its structure, location and high binding affinity to the capsid indicate that the gp10 dimer functions as a molecular staple between neighbouring capsomeres to ensure the particle's stability. Beyond epsilon15, this method potentially offers a new approach for modelling the backbone conformations of the protein subunits in other macromolecular assemblies at near-native solution states.     

The genome of the choanoflagellate Monosiga brevicollis and the origin of metazoans

Choanoflagellates are the closest known relatives of metazoans. To discover potential molecular mechanisms underlying the evolution of metazoan multicellularity, we sequenced and analysed the genome of the unicellular choanoflagellate Monosiga brevicollis. The genome contains approximately 9,200 intron-rich genes, including a number that encode cell adhesion and signalling protein domains that are otherwise restricted to metazoans. Here we show that the physical linkages among protein domains often differ between M. brevicollis and metazoans, suggesting that abundant domain shuffling followed the separation of the choanoflagellate and metazoan lineages. The completion of the M. brevicollis genome allows us to reconstruct with increasing resolution the genomic changes that accompanied the origin of metazoans.     

Evolutionary origin and development of snake fangs

Many advanced snakes use fangs-specialized teeth associated with a venom gland-to introduce venom into prey or attacker. Various front- and rear-fanged groups are recognized, according to whether their fangs are positioned anterior (for example cobras and vipers) or posterior (for example grass snakes) in the upper jaw. A fundamental controversy in snake evolution is whether or not front and rear fangs share the same evolutionary and developmental origin. Resolving this controversy could identify a major evolutionary transition underlying the massive radiation of advanced snakes, and the associated developmental events. Here we examine this issue by visualizing the tooth-forming epithelium in the upper jaw of 96 snake embryos, covering eight species. We use the sonic hedgehog gene as a marker, and three-dimensionally reconstruct the development in 41 of the embryos. We show that front fangs develop from the posterior end of the upper jaw, and are strikingly similar in morphogenesis to rear fangs. This is consistent with their being homologous. In front-fanged snakes, the anterior part of the upper jaw lacks sonic hedgehog expression, and ontogenetic allometry displaces the fang from its posterior developmental origin to its adult front position-consistent with an ancestral posterior position of the front fang. In rear-fanged snakes, the fangs develop from an independent posterior dental lamina and retain their posterior position. In light of our findings, we put forward a new model for the evolution of snake fangs: a posterior subregion of the tooth-forming epithelium became developmentally uncoupled from the remaining dentition, which allowed the posterior teeth to evolve independently and in close association with the venom gland, becoming highly modified in different lineages. This developmental event could have facilitated the massive radiation of advanced snakes in the Cenozoic era, resulting in the spectacular diversity of snakes seen today.