A computer program for comparative analysis of nucleic acid sequences.

The programs offer the possibility of comparing pairs of homologous sequences in order to find out percentage of homology, number of identical and deviating nucleotides, of transitions and transversions and, derived from these, KNUC-values according to Kimura (1) and the corresponding standard error sigmaK. The sequences can be printed in pairs underneath each other, homologies are indicated by asterisks between the identical nucleotides. Out of a set of homologous sequences stored on a disk any number of sequences can be compared in pairs in this way, and a matrix containing either the percentage of homology values, the number of deviating nucleotides or the KNUC-values together with the corresponding standard errors can be sent to screen, printer or disk. A program will be available soon which creates a dendrogram representing the similarity between the sequences by use of an average linkage clustering method deduced from this matrix. The programs are written for Apple II computers using UCSD-PASCAL and for Sirius I/Victor 9000 computers using TURBO-PASCAL.     

De novo design of sequences for nucleic acid structural engineering

An interactive procedure has been developed to assign sequences for the design of nucleic acid secondary structure. The primary goal of the procedure is to facilitate macromolecular architecture studies through the design of branched nucleic acid mono- and oligo-junction constructs in a convenient fashion. The essential feature of the sequence-symmetry minimization algorithm employed is the treatment of short sequences as vocabulary elements whose repetition decreases control over the resulting secondary structure. Both manual and semi-automatic application of this approach are available. The design of linear nucleic acid molecules or molecules containing single-stranded loops or connectors is also possible through application of the procedure.     

A microcomputer program for analysis of nucleic acid hybridization data

The study of nucleic acid hybridization is facilitated by computer mediated fitting of theoretical models to experimental data. This paper describes a non-linear curve fitting program, using the `Patternsearch' algorithm, written in BASIC for the Apple II microcomputer. The advantages and disadvantages of using a microcomputer for local data processing are discussed.     

Comparative analysis of nucleic acid sequences by their general constraints.

We describe two measures of a nucleic acid sequence, derived from Information Theory, which characterize the constraints toward nonuniform base composition, and the constraints on the ordering of the bases. These two measures distinguish extra-chromosomal coding sequences from all other coding sequences examined. The two measures separate eukaryotic coding sequences into two groups: those with introns and those without introns. We have also found a relationship between the general constraints of a subsequence and its degree of conservation in related genes.     

A conversational system for the computer analysis of nucleic acid sequences.

We present a conversational system for the computer analysis of nucleic acid and protein sequences based on the well-known Queen and Korn program (1). The system can be used by persons with only minimal knowledge of computers.     

A rate-independent technique for analysis of nucleic acid sequences: evolutionary parsimony

The method of evolutionary parsimony--or operator invariants--is a technique of nucleic acid sequence analysis related to parsimony analysis and explicitly designed for determining evolutionary relationships among four distantly related taxa. The method is independent of substitution rates because it is derived from consideration of the group properties of substitution operators rather than from an analysis of the probabilities of substitution in branches of a tree. In both parsimony and evolutionary parsimony, three patterns of nucleotide substitution are associated one-to-one with the three topologically linked trees for four taxa. In evolutionary parsimony, the three quantities are operator invariants. These invariants are the remnants of substitutions that have occurred in the interior branch of the tree and are analogous to the substitutions assigned to the central branch by parsimony. The two invariants associated with the incorrect trees must equal zero (statistically), whereas only the correct tree can have a nonzero invariant. The chi 2-test is used to ascertain the nonzero invariant and the statistically favored tree. Examples, obtained using data calculated with evolutionary rates and branchings designed to camouflage the true tree, show that the method accurately predicts the tree, even when substitution rates differ greatly in neighboring peripheral branches (conditions under which parsimony will consistently fail). As the number of substitutions in peripheral branches becomes fewer, the parsimony and the evolutionary-parsimony solutions converge. The method is robust and easy to use.      

A computer program for the estimation of protein and nucleic acid sequence diversity in random point mutagenesis libraries

A computer program for the generation and analysis of in silico random point mutagenesis libraries is described. The program operates by mutagenizing an input nucleic acid sequence according to mutation parameters specified by the user for each sequence position and type of point mutation. The program can mimic almost any type of random mutagenesis library, including those produced via error-prone PCR (ep-PCR), mutator Escherichia coli strains, chemical mutagenesis, and doped or random oligonucleotide synthesis. The program analyzes the generated nucleic acid sequences and/or the associated protein library to produce several estimates of library diversity (number of unique sequences, point mutations, and single point mutants) and the rate of saturation of these diversities during experimental screening or selection of clones. This information allows one to select the optimal screen size for a given mutagenesis library, necessary to efficiently obtain a certain coverage of the sequence-space. The program also reports the abundance of each specific protein mutation at each sequence position, which is useful as a measure of the level and type of mutation bias in the library. Alternatively, one can use the program to evaluate the relative merits of preexisting libraries, or to examine various hypothetical mutation schemes to determine the optimal method for creating a library that serves the screen/selection of interest. Simulated libraries of at least 10⁹ sequences are accessible by the numerical algorithm with currently available personal computers; an analytical algorithm is also available which can rapidly calculate a subset of the numerical statistics in libraries of arbitrarily large size. A multi-type double-strand stochastic model of ep-PCR is developed in an appendix to demonstrate the applicability of the algorithm to amplifying mutagenesis procedures. Estimators of DNA polymerase mutation-type-specific error rates are derived using the model. Analyses of an alpha-synuclein ep-PCR library and NNS synthetic oligonucleotide libraries are given as examples.     

Energy and structural analysis of double nucleic acid triplets

In order to investigate the energy and structural character of RNA-RNA triplets and RNA-DNA duplex base triplets, 64 sets of three-dimensional models of RNA-DNA duplex base triplets and mRNA-tRNA triplex base triplets were constructed and optimized by homologous modeling method using the software InsightII. The comparative statistical method and cluster analysis were adopted to study these features. The result showed: (i) all energy parameters of monomer RNA-DNA hybrid triplets and ternary complexes appeared significantly different; and some parameters related with overall molecules such as overall energy, bond energy and coulomb energy have statistically significant correlations between the structures in vacuum and aquatic solutions while other parameters, including theta energy, phi energy, hydrogen bond energy and non-bond energy, changed significantly, but not continuously. (ii) However, the case of mRNA-tRNA triplets was much more complicated in that only the bond energy's correlation coefficient is -0.8. Typically, the main contribution of GC pairs and G/A/U bases were interesting. The models of RNA-DNA hybrid triplets and mRNA-tRNA triplet should be helpful for the study of base pairing in codons and the biological effectiveness of antisense nucleic acids.     

A versatile digital computer program for non-linear regression analysis

1.

1. A general purpose digital computer program is described for application to biological experiments that require a non-linear regression analysis. The mathematical function, or model, to be fitted to a given set of experimental data is written as a section within the program. Given initial estimates for the parameters of the function, the program uses an iterative procedure to adjust the parameters until the sum of squares of residuals has converged to a minimum.     

GEOSEQ: A Pascal program to calculate statistical geometry parameters of aligned nucleic acid sequences

Statistical geometry in sequence space is a statistical methodused mainly to determine the topology of the divergence (i.e.tree, bundle or net) of a set of aligned sequences by combininghorizontal and vertical positional information. GEOSEQ is adocumented program written in Pascal that calculates the statisticalgeometry parameters necessary to choose between phylogenetictopologies. The input file is an optimal alignment of nucleicacid sequences in PHYLIP format (v. 3.3) with a first line containinginformation regarding sequences as well as some options. Inorder to check the randomization level associated with the obtainedtopology, the program has been implemented with a random generatorof sequences under specified set conditions.     

The number of repeats expected in random nucleic acid sequences and found in genes

Many nucleic acid sequences contain local repeats. These are often considered as traces of evolutionary events such as gene duplications. However, every random sequence of four characters contains a rather large amount of chance repeats. To assess the significance of repeats found in a gene it is necessary to know how large a background of chance repeats has to be expected. Equations are derived that allow the computation of the number of repeats of different lengths and frequencies expected in any random sequence of known chain length and base composition. Tandem repeats are considered as well as repeats interspersed with other sequences. Sample calculations on viral, messenger, ribosomal, and transfer RNA sequences show that some contain no more than the expected background of random repeats, whereas others contain an excess. In the latter case, the distribution of distances between the repeats, as well as their number, can give clues as to the evolutionary events that may have produced them.     

Optimization of nucleic acid sequences

Base sequence influences the structure, mechanics, dynamics, and interactions of nucleic acids. However, studying all possible sequences for a given fragment leads to a number of base combinations that increases exponentially with length. We present here a novel methodology based on a multi-copy approach enabling us to determine which base sequence favors a given structural change or interaction via a single energy minimization. This methodology, termed ADAPT, has been implemented starting from the JUMNA molecular mechanics program by adding special nucleotides, "lexides," containing all four bases, whose contribution to the energy of the system is weighted by continuously variable coefficients. We illustrate the application of this approach in the case of double-stranded DNA by determining the optimal sequences satisfying structural (B-Z transition), mechanical (intrinsic curvature), and interaction (ligand-binding) properties.     

A proposal on metrics for identification using nucleic acid sequences

Abstract The need is stressed for attempts to be made to permit diagnostic nucleic acid sequences to be used in a quantitative manner. Sequence differences or binding values should be converted to a distance measure and from this an ultrametric tree should be constructed. A single quantitative determination can yield considerable information about the likely identity of an unknown microorganism when the distance obtained from the sequence is compared with the tree. The concept is illustrated by hypothetical species and genus subsequences, and it is suitable both for successive use of hierarchical subsequences and for automated identification. It is pointed out that entirely specific subsequences for higher taxa may be difficult to discover. These principles will be useful for the future design of diagnostic sequences, including possible application to DNA-DNA pairing.     

A rapid method of searching for homology of nucleic acid sequences

A new method of the homology search between DNA sequences was suggested. This method may be used to find extensive and not strong homologies with point mutations and deletions. The computer time to compare sequences is less than dynamic program algorithms at least by four orders of magnitude. It makes possible to use the method for homology search all over the nucleotide bank by personal computers. Some results of homology search are presented.     

'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers

DNA Strider is a new integrated DNA and Protein sequence analysis program written with the C language for the Macintosh Plus, SE and II computers. It has been designed as an easy to learn and use program as well as a fast and efficient tool for the day-to-day sequence analysis work. The program consists of a multi-window sequence editor and of various DNA and Protein analysis functions. The editor may use 4 different types of sequences (DNA, degenerate DNA, RNA and one-letter coded protein) and can handle simultaneously 6 sequences of any type up to 32.5 kB each. Negative numbering of the bases is allowed for DNA sequences. All classical restriction and translation analysis functions are present and can be performed in any order on any open sequence or part of a sequence. The main feature of the program is that the same analysis function can be repeated several times on different sequences, thus generating multiple windows on the screen. Many graphic capabilities have been incorporated such as graphic restriction map, hydrophobicity profile and the CAI plot- codon adaptation index according to Sharp and Li. The restriction sites search uses a newly designed fast hexamer look-ahead algorithm. Typical runtime for the search of all sites with a library of 130 restriction endonucleases is 1 second per 10,000 bases. The circular graphic restriction map of the pBR322 plasmid can be therefore computed from its sequence and displayed on the Macintosh Plus screen within 2 seconds and its multiline restriction map obtained in a scrolling window within 5 seconds.     

Protein structural models for nucleic acid interactions

It is proposed that particular segments of some ribosomal, histone and plant viral capsid proteins adopt a helical structural mode for interaction with nucleic acid. The amino acid regions were determined by three probes applied to 26 protein sequences: searches for helical wheels displaying asymmetric basic charge distributions, secondary structural predictions, and searches for primary structural homologies. In 11 of the protein sequences examined, homologous heptapeptides were found in the residue spans delineated by the three probes. A helical wheel analysis of the oligopeptide amino acids showed a distinct positive charge clustering. It is suggested that the basic amino acid side chains on the hydrophilic helical side interact with nucleic acid negative phosphate groups while the somewhat hydrophobic side is available for interaction within the protein or possibly with the major groove of double-stranded nucleic acid.