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Alanko, J.

Publications and source records attributed to Alanko, J..

3 recordsLinked to original sources

A framework for space-efficient variable-order Markov models

MotivationMarkov models with contexts of variable length are widely used in bioinformatics for representing sets of sequences with similar biological properties. When models contain many long contexts, existing implementations are either unable to handle genome-scale training datasets within typical memory budgets, or they are optimized for specific model variants and are thus inflexible.\n\nResultsWe provide practical, versatile representations of variable-order Markov models and of interpolated Markov models, that support a large number of context-selection criteria, scoring functions, probability smoothing methods, and interpolations, and that take up to 4 times less space than previous implementations based on the suffix array, regardless of the number and length of contexts, and up to 10 times less space than previous trie-based representations, or more, while matching the size of related, state-of-the-art data structures from Natural Language Processing. We describe how to further compress our indexes to a quantity related to the redundancy of the training data, saving up to 90% of their space on repetitive datasets, and making them become up to 60 times smaller than previous implementations based on the suffix array. Finally, we show how to exploit constraints on the length and frequency of contexts to further shrink our compressed indexes to half of their size or more, achieving data structures that are 100 times smaller than previous implementations based on the suffix array, or more. This allows variable-order Markov models to be trained on bigger datasets and with longer contexts on the same hardware, thus possibly enabling new applications.\n\nAvailability and implementationhttps://github.com/jnalanko/VOMM

bioinformatics

GGA2 and RAB13 regulate activity-dependent β1-integrin recycling

{beta}1-integrins mediate cell-matrix interactions and their trafficking is important in the dynamic regulation of cell adhesion, migration and malignant processes like cancer cell invasion. Here we employ an RNAi screen to characterize regulators of integrin traffic and identify the association of Golgi-localized gamma ear-containing Arf-binding protein 2 (GGA2) with {beta}1-integrin and its role in recycling of the active but not inactive {beta}1-integrin receptors. Silencing of GGA2 limits active {beta}1-integrin levels in focal adhesions and decreases cancer cell migration and invasion congruent with its ability to regulate the dynamics of active integrins. Using the proximity-dependent biotin identification (BioID) method, we identify two RAB family small GTPases, RAB13 and RAB10, associating with GGA2 and {beta}1-integrin. Functionally, RAB13 silencing triggers the intracellular accumulation of active {beta}1-integrin, reduces integrin activity, in focal adhesions, and cell migration, similarly to GGA2 depletion, indicating that both facilitate active {beta}1-integrin recycling the plasma membrane. Thus, GGA2 and RAB13 are important specificity determinants for integrin activity-dependent traffic.

cell biology

ProLIF: a quantitative assay for investigating integrin cytoplasmic protein interactions and synergistic membrane effects on proteoliposomes

Integrin transmembrane heterodimeric receptors control a wide range of biological interactions by triggering the assembly of large multiprotein complexes at their cytoplasmic interface. A diverse set of methods have been used to investigate cytoplasmic interactions between integrins and intracellular proteins. These predominantly consist of peptide-based pull-downs and biochemical immuno-isolations from detergent-solubilized cell lysates. However, quantitative methods to probe integrin-protein interactions in a more biologically relevant context where the integrin is embedded within a lipid bilayer have been lacking. Here we describe a technique called ProLIF (Protein-Liposome Iinenteractions by Flow cytometry) to reconstitute recombinant integrin transmembrane domain (TMD) and cytoplasmic tail (CT) fragments on liposomes as individual or {beta} subunits or as {beta} heterodimers and, using flow cytometry, to rapidly and quantitatively measure protein interactions with these membrane-embedded integrins. Importantly, the assay can analyse binding of fluorescent proteins directly from cell lysates without further purification steps. By combining integrins with membrane lipids to generate proteoliposomes, the effects of membrane composition such as PI(4,5)P2 presence on protein recruitment to the integrin CTs can be analyzed. ProLIF requires no specific instrumentation, apart from a standard flow cytometer and can be applied to measure a broad range of membrane-dependent protein-protein interactions with the potential for high-throughput/multiplex analyses.

molecular biology