The Duplexing of the Genetic Code and Sequence-Dependent DNA Geometry

It is well known that sequences of bases in DNA are translated into sequences of amino acids in cells via the genetic code. More recently, it has been discovered that the sequence of DNA bases also influences the geometry and deformability of the DNA. These two correspondences represent a naturally arising example of duplexed codes, providing two different ways of interpreting the same DNA sequence. This paper will set up the notation and basic results necessary to mathematically investigate the relationship between these two natural DNA codes. It then undertakes two very different such investigations: one graphical approach based only on expected values and another analytic approach incorporating the deformability of the DNA molecule and approximating the mutual information of the two codes. Special emphasis is paid to whether there is evidence that pressure to maximize the duplexing efficiency influenced the evolution of the genetic code. Disappointingly, the results fail to support the hypothesis that the genetic code was influenced in this way. In fact, applying both methods to samples of realistic alternative genetic codes shows that the duplexing of the genetic code found in nature is just slightly less efficient than average. The implications of this negative result are considered in the final section of the paper.     

Localization and organization of phenol degradation genes ofPseudomonas putida strain H

The genetic organization of the DNA region encoding the phenol degradation pathway ofPseudomonas putida H has been investigated. This strain can utilize phenol or some of its methylated derivatives as its sole source of carbon and energy. The first step in this process is the conversion of phenol into catechol. Catechol is then further metabolized via themeta-cleavage pathway into TCA cycle intermediates. Genes encoding these enzymes are clustered on the plasmid pPGH1. A region of contiguous DNA spanning about 16 kb contains all of the genetic information necessary for inducible phenol degradation. The analysis of mutants generated by insertion of transposons and cassettes indicates that all of the catabolic genes are contained in a single operon. This codes for a multicomponent phenol hydroxylase andmeta-cleavage pathway enzymes. Catabolic genes are subject to positive control by the gene product(s) of a second locus.     

Validation of an entirely in vitro approach for rapid prototyping of DNA regulatory elements for synthetic biology

A bottleneck in our capacity to rationally and predictably engineer biological systems is the limited number of well-characterized genetic elements from which to build. Current characterization methods are tied to measurements in living systems, the transformation and culturing of which are inherently time-consuming. To address this, we have validated a completely in vitro approach for the characterization of DNA regulatory elements using Escherichia coli extract cell-free systems. Importantly, we demonstrate that characterization in cell-free systems correlates and is reflective of performance in vivo for the most frequently used DNA regulatory elements. Moreover, we devise a rapid and completely in vitro method to generate DNA templates for cell-free systems, bypassing the need for DNA template generation and amplification from living cells. This in vitro approach is significantly quicker than current characterization methods and is amenable to high-throughput techniques, providing a valuable tool for rapidly prototyping libraries of DNA regulatory elements for synthetic biology.     

An information theory of the genetic code

An information theory of the genetic code is given, which deals with the process by which template codes (nucleotides or codons) choose substrate codes (nucleotides or anti-codons) in accordance with the base-pairing rules in the chain elongation phase of polynucleotide or polypeptide synthesis. A definite period of recognition time (τ) required for a template code to discriminate a substrate code is proposed, and an experimental method for determining the time is suggested. A substrate word is defined to be the sequence of substrate codes which have appeared at a recognition site in turn before a substrate code complementary to a template code first appears, and the mean length of substrate words (F) is derived from the mole fractions of template codes and substrate codes. The chain elongation rate is greatest when the mole fractions of template codes is proportional to the square of those of substrate codes to minimize the mean recognition time per word (Fτ). The uncertainty of a template (G) and the uncertainty of a medium (M) respectively are derived from the minimum of the function F. The amount of genetic information contained in a template is measured by the function G. The unit of the amount of genetic information is termed “cit”. The function M, the ratio of the number of all binary collisions to the number of homogeneous binary collisions in a mixture of different molecules, may be the new other “entropy” which represents informational properties of the mixture not represented by thermodynamic entropy of mixing. Both functions (G and M) have maxima when all random variables are equal and they are multiplicative in nature in contrast to entropy which is additive. The multiplicativity of the function G may contribute to the enormous informational capacity of genes.     

DR-GAS: A database of functional genetic variants and their phosphorylation states in human DNA repair systems

We present DR-GAS¹, a unique, consolidated and comprehensive DNA repair genetic association studies database of human DNA repair system. It presents information on repair genes, assorted mechanisms of DNA repair, linkage disequilibrium, haplotype blocks, nsSNPs, phosphorylation sites, associated diseases, and pathways involved in repair systems. DNA repair is an intricate process which plays an essential role in maintaining the integrity of the genome by eradicating the damaging effect of internal and external changes in the genome. Hence, it is crucial to extensively understand the intact process of DNA repair, genes involved, non-synonymous SNPs which perhaps affect the function, phosphorylated residues and other related genetic parameters. All the corresponding entries for DNA repair genes, such as proteins, OMIM IDs, literature references and pathways are cross-referenced to their respective primary databases. DNA repair genes and their associated parameters are either represented in tabular or in graphical form through images elucidated by computational and statistical analyses. It is believed that the database will assist molecular biologists, biotechnologists, therapeutic developers and other scientific community to encounter biologically meaningful information, and meticulous contribution of genetic level information towards treacherous diseases in human DNA repair systems. DR-GAS is freely available for academic and research purposes at: http://www.bioinfoindia.org/drgas.     

Whole-genome sequence diversity and association analysis of 198 soybean accessions in mini-core collections

We performed whole-genome Illumina resequencing of 198 accessions to examine the genetic diversity and facilitate the use of soybean genetic resources and identified 10 million single nucleotide polymorphisms and 2.8 million small indels. Furthermore, PacBio resequencing of 10 accessions was performed, and a total of 2,033 structure variants were identified. Genetic diversity and structure analysis congregated the 198 accessions into three subgroups (Primitive, World, and Japan) and showed the possibility of a long and relatively isolated history of cultivated soybean in Japan. Additionally, the skewed regional distribution of variants in the genome, such as higher structural variations on the R gene clusters in the Japan group, suggested the possibility of selective sweeps during domestication or breeding. A genome-wide association study identified both known and novel causal variants on the genes controlling the flowering period. Novel candidate causal variants were also found on genes related to the seed coat colour by aligning together with Illumina and PacBio reads. The genomic sequences and variants obtained in this study have immense potential to provide information for soybean breeding and genetic studies that may uncover novel alleles or genes involved in agronomically important traits.     

Rule-Based Models of the Interplay between Genetic and Environmental Factors in Childhood Allergy

Both genetic and environmental factors are important for the development of allergic diseases. However, a detailed understanding of how such factors act together is lacking. To elucidate the interplay between genetic and environmental factors in allergic diseases, we used a novel bioinformatics approach that combines feature selection and machine learning. In two materials, PARSIFAL (a European cross-sectional study of 3113 children) and BAMSE (a Swedish birth-cohort including 2033 children), genetic variants as well as environmental and lifestyle factors were evaluated for their contribution to allergic phenotypes. Monte Carlo feature selection and rule based models were used to identify and rank rules describing how combinations of genetic and environmental factors affect the risk of allergic diseases. Novel interactions between genes were suggested and replicated, such as between ORMDL3 and RORA, where certain genotype combinations gave odds ratios for current asthma of 2.1 (95% CI 1.2-3.6) and 3.2 (95% CI 2.0-5.0) in the BAMSE and PARSIFAL children, respectively. Several combinations of environmental factors appeared to be important for the development of allergic disease in children. For example, use of baby formula and antibiotics early in life was associated with an odds ratio of 7.4 (95% CI 4.5-12.0) of developing asthma. Furthermore, genetic variants together with environmental factors seemed to play a role for allergic diseases, such as the use of antibiotics early in life and COL29A1 variants for asthma, and farm living and NPSR1 variants for allergic eczema. Overall, combinations of environmental and life style factors appeared more frequently in the models than combinations solely involving genes. In conclusion, a new bioinformatics approach is described for analyzing complex data, including extensive genetic and environmental information. Interactions identified with this approach could provide useful hints for further in-depth studies of etiological mechanisms and may also strengthen the basis for risk assessment and prevention.     

A Novel Approach for Identifying Causal Models of Complex Diseases from Family Data

Causal models including genetic factors are important for understanding the presentation mechanisms of complex diseases. Familial aggregation and segregation analyses based on polygenic threshold models have been the primary approach to fitting genetic models to the family data of complex diseases. In the current study, an advanced approach to obtaining appropriate causal models for complex diseases based on the sufficient component cause (SCC) model involving combinations of traditional genetics principles was proposed. The probabilities for the entire population, i.e., normal–normal, normal–disease, and disease–disease, were considered for each model for the appropriate handling of common complex diseases. The causal model in the current study included the genetic effects from single genes involving epistasis, complementary gene interactions, gene–environment interactions, and environmental effects. Bayesian inference using a Markov chain Monte Carlo algorithm (MCMC) was used to assess of the proportions of each component for a given population lifetime incidence. This approach is flexible, allowing both common and rare variants within a gene and across multiple genes. An application to schizophrenia data confirmed the complexity of the causal factors. An analysis of diabetes data demonstrated that environmental factors and gene–environment interactions are the main causal factors for type II diabetes. The proposed method is effective and useful for identifying causal models, which can accelerate the development of efficient strategies for identifying causal factors of complex diseases.     

Genetic variation of coreoleuciscus splendidus populations (Teleostei; Cypriniformes) from four major river drainage systems in South Korea as assessed by AFLP markers

Amplified fragment length polymorphism (AFLP) technique was used to assess genetic variation and population structuring of Coreoleuciscus splendidus (Teleostei; Cypriniformes) from four major river drainage systems in South Korea (i.e. the Han and Geum Rivers belonging to the West Korea Subdistrict and the Seomjin and Nakdong Rivers belonging to the South Korea Subdistrict). Four combinations of selective primers generated 152–186 DNA fragments, of which 45–59 were polymorphic. The Geum River population had the lowest genetic variation in terms of average heterozygosity. The pairwise genetic differentiation (F _ST) estimates showed that the four populations of C. splendidus were genetically differentiated (P < 0.05). The F _ST values were higher between population groups of the West and South Subdistricts than between populations from the two river systems within each subdistrict. A dendrogram based on the distance matrix revealed that all C. splendidus individuals bifurcated into two subdistrict population groups and were further divided into two river populations within each subdistrict. Individuals from each population consistently clustered together with high statistical support. The analysis of molecular variance after partitioning of C. splendidus populations into two groups (i.e. the West and South Korea Subdistricts) revealed very strong geographic structuring between them with 74.01% of total variance (P < 0.001). Taken together, the AFLP markers clearly distinguished not only between two population groups of the West and South Subdistricts but also between populations from two river systems within each subdistrict. Morphological differences between the two subdistrict population groups in body color and the presence of black array(s) on dorsal, anal, and caudal fin rays agreed with their high degree of genetic variation. Our findings strongly indicate that restricted dispersal and gene flow are apparent among four geographically separate populations of C. splendidus, and each population should be treated as an evolutionarily significant unit; this should be an a priori consideration for developing their conservation strategies.     

Recent advances in studies of evolutionary relationships between proteins and nucleic acids

Evolution depends upon the occurrence of occasional changes, large or small, in hereditary characteristics. Molecular genetics gave rise to the new field of molecular evolution, which is currently exploring the changes that take place in proteins and nucleic acids over long periods of time. The following are some of the fundamental assumptions:

1. The phenotypic characteristics of organisms depend directly on proteins.
2. Proteins are synthesized in accordance with information carried in molecules of DNA as sequences of the four bases, adenine, guanine, cytosine, and thymine. The information is transcribed into molecules of messenger RNA and is translated into proteins by the intervention of the genetic code.
3. Changes in the composition of the base sequences in DNA can take place in living organisms, and these changes can affect the phenotypic characteristics of the next generation.
4. The process of natural selection favors the perpetuation of organisms which compete successfully in the struggle for existence. This process leads to the elimination of all but a small fraction of the astronomical number of possible protein molecules that could result from genetic translation of the possible variants of DNA. Furthermore, the number of protein molecules was originally much smaller than it is to-day, and it has increased by hereditary processes rather than by the chance appearance of entirely new proteins.
5. The DNA present in any single cell contains the complete information for all the hereditary characteristics of the organism. The amount of DNA per cell may increase during evolution and this increase has produced modern organisms that are ‘higher’, more specialized, and more complex, from carlicr and simpler forms.
6. Protein molecules are slowly and steadily differentiated during evolution if their genes are physically separated from each other, by allopatric speciation or even by duplication and translocation, whether or not the function of the proteins are changed.
7. Mutations, together with recombination, contribute to changes in the genetic pool which provide the variability within populations that is necessary for evolution of species.

The field of molecular evolution should include a theory of the chemical events leading to the formation of the first living organism from molecules of non-living origin. The gentic code may have evolved through multiplication of transfer RNA molecules by gene duplication followed by differentiation. This proposal is supported by the similarities between all tRNA molecules of known structures. The DNA of higher organisms contains families of repetitive sequences. The families may contain thousands or hundreds of thousands of individual members. The ‘family resemblance’ within each group grows less with the passage of time because this leads to differentiation resulting from the accumulation of point mutations.     

Biological pathways to adaptability--interactions between genome, epigenome, nervous system and environment for adaptive behavior

Because living systems depend on their environment, the evolution of environmental adaptability is inseparable from the evolution of life itself (Pross 2003). In animals and humans, environmental adaptability extends further to adaptive behavior. It has recently emerged that individual adaptability depends on the interaction of adaptation mechanisms at diverse functional levels. This interaction enables the integration of genetic, epigenetic and environmental factors for coordinated regulation of adaptations. In this review, we first present the basis for the regulation of adaptation mechanisms across functional levels. We then focus on neuronal activity-regulated adaptation mechanisms that involve the regulation of genes, noncoding DNA (ncDNA), ncRNAs and proteins to change the structural and functional properties of neurons. Finally, we discuss a selection of these important neuronal activity-regulated molecules and their effects on brain structure and function and on behavior. Most of the evidence so far is based on sampling of animal tissue or post-mortem studies in humans. However, we also present techniques that combine genetic with behavioral and neurophysiological measures in humans (e.g. genetic imaging) and discuss their potential and limitations. We argue that we need to understand how neuronal activity-dependent adaptation mechanisms integrate genetic, epigenetic and experience-dependent signals in order to explain individual variations in behavior and cognitive performance.     

On the information content of the genetic code

In living organisms 20 amino acids along with the terminator value(s) are encoded by 64 codons giving a degeneracy of the codons as described by the genetic code. A basic theoretical problem of genetic codes is to explain the particular distribution of degeneracies of partitions involved in the codes. In this work the degeneracy problem is considered in the framework of information theory. It is shown by direct numerical evaluation of a certain degeneracy information function associated with the genetic code that the degeneracy of the codes is observed to be related to the optimization of this function.     

The Escherichia coli SOS gene dinF protects against oxidative stress and bile salts

DNA is constantly damaged by physical and chemical factors, including reactive oxygen species (ROS), such as superoxide radical (O₂ ⁻), hydrogen peroxide (H₂O₂) and hydroxyl radical (•OH). Specific mechanisms to protect and repair DNA lesions produced by ROS have been developed in living beings. In Escherichia coli the SOS system, an inducible response activated to rescue cells from severe DNA damage, is a network that regulates the expression of more than 40 genes in response to this damage, many of them playing important roles in DNA damage tolerance mechanisms. Although the function of most of these genes has been elucidated, the activity of some others, such as dinF, remains unknown. The DinF deduced polypeptide sequence shows a high homology with membrane proteins of the multidrug and toxic compound extrusion (MATE) family. We describe here that expression of dinF protects against bile salts, probably by decreasing the effects of ROS, which is consistent with the observed decrease in H₂O₂-killing and protein carbonylation. These results, together with its ability to decrease the level of intracellular ROS, suggests that DinF can detoxify, either direct or indirectly, oxidizing molecules that can damage DNA and proteins from both the bacterial metabolism and the environment. Although the exact mechanism of DinF activity remains to be identified, we describe for the first time a role for dinF.     

The global bacterial regulator H-NS promotes transpososome formation and transposition in the Tn5 system

The histone-like nucleoid structuring protein (H-NS) is an important regulator of stress response and virulence genes in gram-negative bacteria. In addition to binding regulatory regions of genes in a structure-specific manner, H-NS also binds in a structure-specific manner to sites in the Tn10 transpososome, allowing it to act as a positive regulator of Tn10 transposition. This is the only example to date of H-NS regulating a transposition system by interacting directly with the transposition machinery. In general, transposition complexes tend to include segments of deformed DNA and given the capacity of H-NS to bind such structures, and the results from the Tn10 system, we asked if H-NS might regulate another transposition system (Tn5) by directly binding the transposition machinery. We show in the current work that H-NS does bind Tn5 transposition complexes and use hydroxyl radical footprinting to characterize the H-NS interaction with the Tn5 transpososome. We also show that H-NS can promote Tn5 transpososome formation in vitro, which correlates with the Tn5 system showing a dependence on H-NS for transposition in vivo. Taken together the results suggest that H-NS might play an important role in the regulation of many different bacterial transposition systems and thereby contribute directly to lateral gene transfer.     

Barbieri’s Organic Codes Enable Error Correction of Genomes

Barbieri introduced and developed the concept of organic codes. The most basic of them is the genetic code, a set of correspondence rules between otherwise unrelated sequences: strings of nucleotides on the one hand, polypeptidic chains on the other hand. Barbieri noticed that it implies ‘coding by convention’ as arbitrary as the semantic relations a language establishes between words and outer objects. Moreover, the major transitions in life evolution originated in new organic codes similarly involving conventional rules. Independently, dealing with heredity as communication over time and relying on information theory, we asserted that the conservation of genomes over the ages demands that error-correcting codes make them resilient to casual errors. Moreover, the better conservation of very old parts of the genome demands that they result from combining successively established nested codes such that the older an information, the more numerous component codes protect it. Barbieri’s concept of organic code and that of genomic error-correcting code may seem unrelated. We show however that organic codes actually entail error-correcting properties. Error-correcting, in general, results from constraints being imposed on a set of sequences. Mathematical equalities are conveniently used in communication engineering for expressing constraints but error correction only needs that constraints exist. Biological sequences are similarly endowed with error-correcting ability by physical-chemical or linguistic constraints, thus defining ‘soft codes’. These constraints are moreover presumably efficient for correcting errors. Insofar as biological sequences are subjected to constraints, organic codes necessarily involve soft codes, and their successive onset results in the nested structure we hypothesized. Organic codes are generated and maintained by means of molecular ‘semantic feedback loops’. Each of these loops involves genes which code for proteins, the enzymatic action of which controls a function needed for the protein assembly. Taken together, thus, they control the assembly of their own structure as instructed by the genome and, once closed, these loops ensure their own conservation. However, the semantic feedback loops do not prevent the genome lengthening. It increases both the redundancy of the genome (as an error-correcting code) and the information quantity it bears, thus improving the genome reliability and the specificity of the enzymes, which enables further evolution.