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em Nottingham eTheses


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This paper reports some experiments in using SVG (Scalable Vector Graphics), rather than the browser default of (X)HTML/CSS, as a potential Web-based rendering technology, in an attempt to create an approach that integrates the structural and display aspects of a Web document in a single XML-compliant envelope. Although the syntax of SVG is XML based, the semantics of the primitive graphic operations more closely resemble those of page description languages such as PostScript or PDF. The principal usage of SVG, so far, is for inserting complex graphic material into Web pages that are predominantly controlled via (X)HTML and CSS. The conversion of structured and unstructured PDF into SVG is discussed. It is found that unstructured PDF converts into pages of SVG with few problems, but difficulties arise when one attempts to map the structural components of a Tagged PDF into an XML skeleton underlying the corresponding SVG. These difficulties are not fundamentally syntactic; they arise largely because browsers are innately bound to (X)HTML/CSS as their default rendering model. Some suggestions are made for ways in which SVG could be more totally integrated into browser functionality, with the possibility that future browsers might be able to use SVG as their default rendering paradigm.

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DnaD is a primosomal protein that remodels supercoiled plasmids. It binds to supercoiled forms and converts them to open forms without nicking. During this remodeling process, all the writhe is converted to twist and the plasmids are held around the periphery of large scaffolds made up of DnaD molecules. This DNA-remodeling function is the sum of a scaffold-forming activity on the N-terminal domain and a DNA-dependent oligomerization activity on the C-terminal domain. We have determined the crystal structure of the scaffold-forming N-terminal domain, which reveals a winged-helix architecture, with additional structural elements extending from both N- and C-termini. Four monomers form dimers that join into a tetramer. The N-terminal extension mediates dimerization and tetramerization, with extensive interactions and distinct interfaces. The wings and helices of the winged-helix domains remain exposed on the surface of the tetramer. Structure-guided mutagenesis and atomic force microscopy imaging indicate that these elements, together with the C-terminal extension, are involved in scaffold formation. Based upon our data, we propose a model for the DnaD-mediated scaffold formation.

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The essential Bacillus subtilis DnaD and DnaB proteins have been implicated in the initiation of DNA replication. Recently, DNA remodeling activities associated with both proteins were discovered that could provide a link between global or local nucleoid remodeling and initiation of replication. DnaD forms scaffolds and opens up supercoiled plasmids without nicking to form open circular complexes, while DnaB acts as a lateral compaction protein. Here we show that DnaD-mediated opening of supercoiled plasmids is accompanied by significant untwisting of DNA. The net result is the conversion of writhe (Wr) into negative twist (Tw), thus maintaining the linking number (Lk) constant. These changes in supercoiling will reduce the considerable energy required to open up closed circular plectonemic DNA and may be significant in the priming of DNA replication. By comparison, DnaB does not affect significantly the supercoiling of plasmids. Binding of the DnaD C-terminal domain (Cd) to DNA is not sufficient to convert Wr into negative Tw, implying that the formation of scaffolds is essential for duplex untwisting. Overall, our data suggest that the topological effects of the two proteins on supercoiled DNA are different; DnaD opens up, untwists and converts plectonemic DNA to a more paranemic form, whereas DnaB does not affect supercoiling significantly and condenses DNA only via its lateral compaction activity. The significance of these findings in the initiation of DNA replication is discussed.