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Mining, compressing and classifying with extensible motifs Abstract Background Motif patterns of maximal saturation emerged originally in contexts of pattern discovery in biomolecular sequences and have recently proven a valuable notion also in the design of data compression schemes. Informally, a motif is a string of intermittently solid and wild characters that recurs more or less frequently in an input sequence or family of sequences. Motif discovery techniques and tools tend to be computationally imposing, however, special classes of "rigid" motifs have been identified of which the discovery is affordable in low polynomial time. Results In the present work, "extensible" motifs are considered such that each sequence of gaps comes endowed with some elasticity, whereby the same pattern may be stretched to fit segments of the source that match all the solid characters but are otherwise of different lengths. A few applications of this notion are then described. In applications of data compression by textual substitution, extensible motifs are seen to bring savings on the size of the codebook, and hence to improve compression. In germane contexts, in which compressibility is used in its dual role as a basis for structural inference and classification, extensible motifs are seen to support unsupervised classification and phylogeny reconstruction. Conclusion Off-line compression based on extensible motifs can be used advantageously to compress and classify biological sequences. Background Let s be a sequence of sets of characters from an alphabet Σ ⋃ {·}, where '.' ∉ Σ denotes a don't care (dot, for short) and the rest are solid characters, we use σ to denote a singleton character. For characters e1 and e2, we write e1 ∘ e2 if and only if e1 is a dot or e1 = e2. Allowing for spacers in a string is what makes it extensible. Such spacers are indicated by annotating the dot characters. Specifically, an annotated "." character is written as .α where α is a set of positive integers {α1, α2, ..., αk} or an interval α = [αl, αu], representing all integers between αl and αu including αl and αu. Whenever defined, d will denote the maximum number of consecutive dots allowed in a string. In such cases, for clarity of notation, we use the extensible wild card denoted by the dash symbol "-" instead of the annotated dot character, .[1,d] in the string. Note that '-' ∉ Σ. Thus a string of the form a.[1,d]b will be simply written as a-b. A motif m is extensible if it contains at least one annotated dot, otherwise m is rigid. Given an extensible string m, a rigid string m' is a realization of m if each annotated dot .α is replaced by l ∈ α dots. The collection of all such rigid realizations of m is denoted by R(m). A rigid string m occurs at position l on s if m[j] ∘ s[l + j - 1] holds for 1 ≤ j ≤ |m|. An extensible string m occurs at position l in s if there exists a realization m' of m that occurs at l. Note than an extensible string m could possibly occur more than once at a location on a sequence s. Throughout in the discussion we are interested mostly in the (unique) first left-most occurrence at each location. For a sequence s and positive integer k, k ≤ |s|, a string (extensible or rigid) m is a motif of s with |m| > 1 and location list m = (l1, l2, ..., lp), if both m[1] and m[|m|] are solid and m, |m| ≥ k, is the list of all and only the occurrences of m in s. Given a motif m let m[j1], m[j2], ... m[jl] be the l solid elements in the motif m. Then the sub-motifs of m are given as follows: for every ji, jt, the sub-motif m[ji ... jt] is obtained by dropping all the elements before (to the left of) ji and all elements after (to the right of) jt in m. We also say that m is a condensation for any of its sub-motifs. We are interested in motifs for which any condensation would disrupt the list of occurrences. Formally, let m1, m2, ..., mj be the motifs in a string s. A motif mi is maximal in length if there exists no ml, l ≠ i with and mi is a sub-motif of ml. A motif mi is maximal in composition if no dot character of mi can be replaced by a solid character that appears in all the locations in m. A motif mi is maximal in extension if no annotated dot character of mi can be replaced by a fixed length substring (without annotated dot characters) that appears in all the locations in m. A maximal motif is maximal in composition, in extension and in length. For an exhaustive description of these properties we refer the reader to [1]. Results and discussion Several measures of distance have been proposed and used to classify documents of diverse nature and to infer relationships among them. In practice, each measure translates in a computational task which might be more or less of a burden. In domains such as genome analysis and natural language processing, the increasing availability of longer and longer sequences and more and more massive data sets is playing havoc with similarity measures based on edit computations and the likes [2]. As an alternative, succinct scores related to compressibility -interpreted as a measure of structural complexity or information contents- have been deployed, of which the lineage may be traced back to Kolmogorov's complexity. The Kolmogorov complexity of a string x, denoted K(x), is the length of the shortest program that would cause a standard universal computer to output x. Along the same lines, the conditional Kolmogorov complexity K(x|y) for strings x and y is defined as the length of the shortest program that, given y as input, will output x as the result. Intuitively, the conditional complexity expresses the information difference between the strings x and y. We refer the reader to, e.g., [3] for a detailed treatment of the theory. Whereas the original Kolmogorov complexity is hardly computable, important emulators have been developed since [4], which conjugate compressibility and ease of computation. Following in these steps, we now test the discriminating power of the data compression method that is based on our Off-line steepest descent paradigm with extensible motifs. In this paper, we present lossy off-line data compression techniques by textual substitution in which the patterns used in compression are chosen among the extensible motifs that are found to recur in the textstring with a minimum pre-specified frequency. Motif discovery and motif-driven parses of various kinds have been previously introduced and used in [5]. Whereas the motifs considered in those studies are "rigid", here we assume that each sequence of gaps present in a motif comes endowed with some individually prescribed degree of elasticity, whereby a same pattern may be stretched to fit segments of the source that match all the solid characters but are otherwise of different lengths. This is expected to save on the size of the codebook, and hence to improve compression. The figure of compression achieved by our algorithm shows good sensitivity in telling apart veritable families of proteins from spurious blends. This sets forth an approach to classification that does away with alignment. The data used for the test consists of protein sequences, which are known to be hardly compressible at all [6]. The experiment reported below uses three different families which were picked at random from the PROSITE repository: AP endonucleases (acnucl), G-protein coupled receptors (gprot) and Succinyl-CoA ligases (succ). Table 2 summarizes the results of lossy and lossless compression for various values of the parameters. The artificial groups are marked "-mix", the last column shows the lossless compression ratio of fake over faithful families, when using motifs with the same parameter values. In all cases, the artificial families show compression ratios that are poorer by 10/20%, and the superiority of the lossy variants manifests itself throughout. The experiments thus verify the discrimination potential of data compression by extensible motifs. It seems thus meaningful to build a classifier on top of this measure. Compressibility by extensible motifs may be used to set up a similarity measure on sequences to be used in the inference of phylogeny. The measure could be extended into a metric distance, along the lines of [7]. Specifically, we denote by Off-line(z) the output size obtained when compressing a string z using the lossless variant of our paradigm, and compute the quantity: Table 1 The pseudocode of the motif extraction algorithm. Main() Iterate(m, B, Result) { { Result ← {}; G:l     m' ← m; B ← {mi|mi is a cell}; G:2     For each b = Extract(B) with For each m = Extract(B) G:3        ((b ~-compatible m' Iterate(m, B, Result);           OR (m' ~-compatible b)) Result ← Result; G:4              If (m' ~-compatible b) } G:5                 mt ← m' ~ b; G:6                 If Nodelnconsistent(mt) exit; G:7                 If (|m'| = |b|) B ← B - {b}; G:8                 If (|| ≤ K) G:9                    m' ← mt; G:10                    Iterate(m', B, Result); G:11              If (b ~-compatible m') G:12                 mt ← b ~ m'; G:13                 If Nodelnconsistent(mt) exit; G:14                 If (|m'| = |b|) B ← B - {b}; G:15                 If (|| ≥ K) G:16                    m' ← mt; G:17                    Iterate(m', B, Result); G:18              For each r ∈ Result with r = m' G:19                 If (m' is not maximal w.r.t. r) return; G:20              Result ← Result ⋃ {m'}; } Table 2 Comparing sensitivity of lossy versus lossless compression by Off-line with Extensible Motifs, as applied to real and fake protein families. File File len param density Lossy Lossless Compr ratio % K D acnucl 4197 10 3 1717 2425 4197 5 3 1757 2370 acnucl-mix 4149 10 3 1864 2629 +8.4 4149 5 3 1972 2621 +10.6 gprot 25482 30 3 7003 9879 25482 20 3 7399 10276 gprot-mix 25335 30 3 8179 11945 +20.9 25335 20 3 8386 11978 +16.5 succ 16297 20 3 4994 7449 16297 10 3 4977 7466 succ-mix 16410 20 3 5929 8803 +18.2 16410 10 3 6415 8962 +19.2 where (xy) denotes the concatenation of x and y. Hence, D(x, y) measures the improvement over Off-line(y) that is brought about by using x as a "dictionary" when compressing y. In the following experiment we construct a phylogeny of the Eutherian orders using complete unaligned mitochondrial genomes of the following 15 mammals from GenBank: human (Homo sapiens [GenBank:V00662]), chimpanzee (Pan troglodytes [GenBank:D38116]), pigmy chimpanzee (Pan paniscus [GenBank:D38113]), gorilla (Gorilla gorilla [GenBank:D38114]), orangutan (Pongo pygmaeus [GenBank:D38115]), gibbon (Hylobates lar [GenBank:X99256]), sumatran orangutan (Pongo pygmaeus abelii [GenBank:X97707]), horse (Equus caballus [GenBank:X79547]), white rhino (Ceratotherium simum [GenBank:Y07726]), harbor seal (Phoca vitulina [GenBank:X63726]), gray seal (Halichoerus grypus [GenBank:X72004]), cat (Felis catus [GenBank:U20753]), finback whale (Balenoptera physalus [GenBank:X61145]), blue whale (Balenoptera musculus [GenBank:X72204]), rat (Rattus norvegicus [GenBank:X14848]) and house mouse (Mus musculus [GenBank:V00711]). The evolutionary tree in Figure 1 is generated by a variant of the classical neighbor-join where instead of minimizing the distances between nodes we maximized the separation. Specifically, for each pair (x, y) of sequences, the quantity D(x, y) is computed. Next, the neighbor-join algorithm is used to build the tree from the matrix of distances. This algorithms selects a pair of (x, y) among those achieving the minimum value for D, and creates an internal node as their father. It then coalesces x and y into a combined sequence the D value of which is computed as the maximum (instead of the average) of those of x and y. The process is continued until the D-matrix has shrunk to a scalar. The first notable finding is that closely related species are indeed grouped together, e.g., grayseal with harboseal, orangutan with sumatranorang, etc. Whereas there is no gold standard for the entire tree, biologists do suggest the following grouping for this case: Figure 1 The evolutionary tree built from complete mammalian mtDNA sequences of 15 species. [width = 450 pt]tree1.eps • Eutheria-Rodens: housemouse, rat. • Primates: chimpa, gibbon, gorilla, human, orangutan, pigmychimpa, sumatranorang. • Ferungulates: bluewhale, finbackwhale, grayseal, harboseal, horse, whiterhino. The phylogeny obtained in our experiment is very close to the commonly accepted ones, which suggests that even a method of compression based on a single type of regularity, as opposed to those that take into account palindromes and other structures may support good comparative genomics. For a comparison, the same treatment was applied to human language text classification, in analogy with what is found in [7,8]. Figure 2 displays the tree obtained in experiments performed with a small subset of languages on the widely translated "Universal Declaration of Human Rights". Once more, the resulting tree is coherent with commonly accepted ones. Figure 2 A partial tree of languages using a distance based on compression by extensible motifs. [width = 280 pt]tree2.eps Conclusion Comparisons of the compression ratios respectively achieved by rigid and extensible motifs displays that the latter bring about additional savings in compression. This suggests that extensible motifs may be preferred to rigid ones also in those cases where they are used as bases for similarity measure and classification among sequences. The unsupervised classification method built on top of such measures have been shown here to consistently produce phylogenic trees for species genomes as well language classifications built on text documents. Methods Mining extensible motifs The procedure of motif extraction that is described in Table 1 essentially constructs the inexact suffix tree of [1] implicitly, in a different order. The input is a string s of size n and two positive integers, K and D. The extensibility parameter D is interpreted in the sense that up to D (or 1 to D) dot characters between two consecutive solid characters are allowed. The output is all maximal extensible (with D spacers) patterns that occur at least K times in s. Incidentally, the algorithm can be adapted to extract rigid motifs as a special case. For this, it suffices to interpret D as the maximum number of dot characters between two consecutive solid characters. The algorithm works by converting the input into a sequence of possibly overlapping cells: A cell is the smallest substring in any pattern on s, that has exactly two solid characters: one at the start and the other at the end position of this substring. A maximal extensible pattern is a sequence of cells. Initialization phase The cell is the smallest extensible component of a maximal pattern and the string can be viewed as a sequence of overlapping cells. If no don't care characters are allowed in the motifs then the cells are non-overlapping. The initialization phase has the following steps. Step 1: Construct patterns that have exactly two solid characters in them and separated by no more than D spaces or "." characters. This is done by scanning the string s from left to right. Further, for each location we store start and end position of the pattern. For example, if s = abzdabyxd and K = 2, D = 2, then all the patterns generated at this step are: ab, a.z, a..d, bz, b.d, b..a, zd, z.a, z..b, da, d.b, d..y, a.y, a..x, by, b.x, b..d, yx, y.d, xd, each with its occurrence list. Thus ab = {(1, 2), (5, 6)}, a.z = {(1, 3)} and so on. Step 2: The extensible cells are constructed by combining all the cells with at least one dot character and the same start and end solid characters. The location list is updated to reflect the start and end position of each occurrence. Continuing the previous example, b-d is generated at this step with b-d = {(2, 4), (6, 9)}. All cells m with |m| 0 dot characters respectively and d1

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