ANalysis of multifunctiOnal genes and Their pAralogs in TEleosts: Identification, Evolution and Annotation through Transcriptome scans and Functional studies
Aim of the present proposal is to identify, characterise and annotate new or known duplicated genes across teleost and non-teleost fish species, to search for their paralogs gene(s) and to examine the possibility of new gene families (if more then one paralogs is found). Therefore a three level approach is suggested. The first level investigates the identification of duplicated genes via transcriptome scans through main marine teleost fish orders and subsequently performs first annotations using BLASTN and BLASTX search against NCBI, Ensemble and other fish specific databases. Further annotation like pattern search, structural characterization and domain detection will be performed. The second level comprises evolutionary studies of the identified genes throughout the selected marine fish species including phylogenetic analysis, synteny analysis, investigations to unravel whether genes under study or gene groups evolve at different rates, gene loss and whether Darwinian selection has occurred. The third level concerns functional studies of the candidate gene set. It is expected to detect diversifications between paralogs genes in terms of expression and evolution. In addition extended research for selected genes will be carried out to determine possible multifunctional properties. Multifunctional properties have been shown for genes which originally were assigned to classical functions like for example Glyceraldehyd-3-phosphate (GAPDH) or warm temperature acclimation protein 65 (WAP65). The former is known as classical housekeeping gene. However divers studies have shown its implementation in development and immune response . Concerning WAP65, it was primarily identified due to increased expression associated with warm temperature acclimation of gold fish and carp [2,3]. In contrast to heat-shock proteins, which were first identified after short-term adjustment, Wap65 was found to be higher expressed due to temperature acclimation (Kikuchi et al., 1993). The term “heat shock protein” was chosen based on early observations on Drosophila exposed to heat stress. Today heat-shock proteins can be up-regulated in cells that are exposed to a broad range of stressors. Similar the term “warm-temperature protein” was assigned to a 65-kDa protein due to its high abundance in various tissues after warm temperate acclimation. At present several studies showed that Wap65 proteins are involved in development, immune response as well as stress response. In the viewpoint of evolution it is of interest to explore whether multifunctionality inhibits adaptability (e.g. by presence or absence of paralogs) or whether organisms profit of multifunctional genes. The outcome of ANnOTATE will highly interest researchers studying gene evolution as well as for researchers interested in candidate genes for biomarker linked to the immune response, egg quality and environmental adaptation. In addition it will contribute significantly to the appropriate annotation of fish genes taking into account the evolutionary relationships of paralogs that have arisen in different vertebrate lineages. The enrichment of the dataset of hagfish as well as for sturgeon is of importance not only to identity the homologues genes which will contribute to phylogenetic analysis and will shed light into the original function of the transcripts but also to the hagfish and sturgeon community. In long-term aspect the outcome may be of importance also in biomedical research as belonging to the vertebrates, several research results in teleosts species have been served as groundwork in human disease research.