Nuria Forns

Single-molecule derivation of salt dependent base-pair free energies in DNA

Accurate knowledge of the thermodynamic properties of nucleic acids is crucial to predicting their structure and stability. To date most measurements of base-pair free energies in DNA are obtained in thermal denaturation experiments, which depend on several assumptions. Here we report measurements of the DNA base-pair free energies based on a simplified system, the mechanical unzipping of single DNA molecules. By combining experimental data with a physical model and an optimization algorithm for analysis, we measure the 10 unique nearest-neighbor base-pair free energies with 0.1 kcal mol-1 precision over two orders of magnitude of monovalent salt concentration. We find an improved set of standard energy values compared with Unified Oligonucleotide energies and a unique set of 10 base-pair-specific salt-correction values. The latter are found to be strongest for AA/TT and weakest for CC/GG. Our unique energy values and salt corrections improve predictions of DNA unzipping forces and are fully compatible with melting temperatures for oligos. The method should make it possible to obtain free energies, enthalpies, and entropies in conditions not accessible by bulk methodologies.

YdgT, the Hha paralogue in Escherichia coli , forms heteromeric complexes with H-NS and StpA

In enteric bacteria, proteins of the Hha/YmoA family play a role in the regulation of gene expression in response to environmental factors. Interaction of both Hha and YmoA with H‐NS has been reported, and an Hha/H‐NS complex has been shown to modulate expression in Escherichia coli of the haemolysin operon of plasmid pHly152. In addition to the hns gene, the chromosome of E. coli and other enteric bacteria also includes the stpA gene that encodes the StpA protein, an H‐NS paralogue. We report here the identification of the Hha paralogue in E. coli, the YdgT protein. As Hha paralogue, YdgT appears to fulfil some of the functions reported for StpA as H‐NS paralogue: YdgT is overexpressed in hha mutants and can compensate, at least partially, some of the hha‐induced phenotypes. We also demonstrate that YdgT interacts both with H‐NS and with StpA. Protein cross‐linking studies showed that YdgT/H‐NS heteromeric complexes are generated within the bacterial cell. The StpA protein, which is subjected to Lon‐mediated turnover, was less stable in the absence of Hha or YdgT. Our findings suggest that Hha, YdgT and StpA may form complexes in vivo.

Temperature-Dependent Conjugative Transfer of R27: Role of Chromosome- and Plasmid-Encoded Hha and H-NS Proteins

IncHI plasmids encode multiple-antibiotic resistance in Salmonella enterica serovar Typhi. These plasmids have been considered to play a relevant role in the persistence and reemergence of this microorganism. The IncHI1 plasmid R27, which can be considered the prototype of IncHI plasmids, is thermosensitive for transfer. Conjugation frequency is highest at low temperature (25 to 30 degrees C), decreasing when temperature increases. R27 codifies an H-NS-like protein (open reading frame 164 [ORF164]) and an Hha-like protein (ORF182). The H-NS and Hha proteins participate in the thermoregulation of gene expression in Escherichia coli. Here we investigated the hypothetical role of such proteins in thermoregulation of R27 conjugation. At a nonpermissive temperature (33 degrees C), transcription of several ORFs in both transfer region 1 (Tra1) and Tra2 from R27 is upregulated in cells depleted of Hha-like and H-NS-like proteins. Both chromosome- and plasmid-encoded Hha and H-NS proteins appear to potentially modulate R27 transfer. The function of R27-encoded Hha-like and H-NS proteins is not restricted to modulation of R27 transfer. Different mutant phenotypes associated with both chromosomal hha and hns mutations are compensated in cells harboring R27.

The challenges of statistical patterns of language: The case of Menzerath's law in genomes

The importance of statistical patterns of language has been debated over decades. Although Zipfs law is perhaps the most popular case, recently, Menzeraths law has begun to be involved. Menzeraths law manifests in language, music and genomes as a tendency of the mean size of the parts to decrease as the number of parts increases in many situations. This statistical regularity emerges also in the context of genomes, for instance, as a tendency of species with more chromosomes to have a smaller mean chromosome size. It has been argued that the instantiation of this law in genomes is not indicative of any parallel between language and genomes because (a) the law is inevitable and (b) non-coding DNA dominates genomes. Here mathematical, statistical and conceptual challenges of these criticisms are discussed. Two major conclusions are drawn: the law is not inevitable and languages also have a correlate of non-coding DNA. However, the wide range of manifestations of the law in and outside genomes suggests that the striking similarities between non-coding DNA and certain linguistics units could be anecdotal for understanding the recurrence of that statistical law.

Size of the Whole versus Number of Parts in Genomes

It is known that chromosome number tends to decrease as genome size increases in angiosperm plants. Here the relationship between number of parts (the chromosomes) and size of the whole (the genome) is studied for other groups of organisms from different kingdoms. Two major results are obtained. First, the finding of relationships of the kind “the more parts the smaller the whole” as in angiosperms, but also relationships of the kind “the more parts the larger the whole”. Second, these dependencies are not linear in general. The implications of the dependencies between genome size and chromosome number are two-fold. First, they indicate that arguments against the relevance of the finding of negative correlations consistent with Menzerath-Altmann law (a linguistic law that relates the size of the parts with the size of the whole) in genomes are seriously flawed. Second, they unravel the weakness of a recent model of chromosome lengths based upon random breakage that assumes that chromosome number and genome size are independent.

Folding and unfolding of a triple-branch DNA molecule with four conformational states

Single-molecule experiments provide new insights into biological processes hitherto not accessible by measurements performed on bulk systems. We report on a study of the kinetics of a triple-branch DNA molecule with four conformational states by pulling experiments with optical tweezers and theoretical modelling. Three distinct force rips associated with different transitions between the conformational states are observed in the folding and unfolding trajectories. By applying transition rate theory to a free energy model of the molecule, probability distributions for the first rupture forces of the different transitions are calculated. Good agreement of the theoretical predictions with the experimental findings is achieved. Furthermore, due to our specific design of the molecule, we found a useful method to identify permanently frayed molecules by estimating the number of opened base-pairs from the measured force jump values.

The Parameters of the Menzerath-Altmann Law in Genomes

Abstract The relationship between the size of the whole and the size of the parts in language and music is known to follow the Menzerath-Altmann law at many levels of description (morphemes, words, sentences, …). Qualitatively, the law states that the larger the whole, the smaller its parts, e.g. the longer a word (in syllables) the shorter its syllables (in letters or phonemes). This patterning has also been found in genomes: the longer a genome (in chromosomes), the shorter its chromosomes (in base pairs). However, it has been argued recently that mean chromosome length is trivially a pure power function of chromosome number with an exponent of −1. The functional dependency between mean chromosome size and chromosome number in groups of organisms from three different kingdoms is studied. The fit of a pure power function yields exponents between −1.6 and 0.1. It is shown that an exponent of −1 is unlikely for fungi, gymnosperm plants, insects, reptiles, ray-finned fishes and amphibians. Even when the exponent is very close to −1, adding an exponential component is able to yield a better fit with regard to a pure power-law in plants, mammals, ray-finned fishes and amphibians. The parameters of the Menzerath-Altmann law in genomes deviate significantly from a power law with a −1 exponent with the exception of birds and cartilaginous fishes.