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Previous studies, the single flagellum of T. brucei changes in morphology and function during the life cycle alternating Sermorelin supplier between a tsetse fly and a mammal host [3,37]. A recent study analyzed the expression profiles of the T. brucei proteome at three important life-cycle stages, namely long slender and short stumpy bloodstream forms in the mammalian host and the procyclic form in the midgut of the tsetse fly [38]. Considering uniquely mapped reads, 800 (83.4 ) flagellar genes were detected to be expressed in at least one of the three stages. Differentially expressed genes were defined to be those two-fold up- or downregulated at two stages with p0:001 according to the Audic and Claverie test [39]. In total, 363 flagellar protein-encoded genes Madrasin significantly changed expression levels in at least one of the three stages, accounting for ,45 of the expressed flagellar proteins (Table 2). As expected, much more flagellar genes changed their expression levels in procyclic form when compared with the other two bloodstream stages. As T. brucei lives in from the tsetse fly to the mammal host, the parasite needs more genes to be regulated to adapt to host change for survival. We found that most of these differentially expressed genes were up-regulated in the procyclic form when compared with the long slender and short stumpybloodstream form. This is not surprising, as we know that the flagellum-mediated migration between the midgut and salivary glands of its tsetse fly vector is essential for the progression of its life cycle [3,40]. When compared with the short stumpy bloodstream form, we found much more flagellar genes were up-regulated in the long slender and procyclic forms. This may due to the important roles of the single flagellum in cell division as demonstrated by previous studies [5,40,41,42], while both the long slender bloodstream form and the procyclic form are proliferative forms. These results indicate life cycle stage-specific regulation of flagellar functions in T. brucei.ConclusionsThe available evidence indicates the multifunctional nature of the single flagellum in T. brucei, and suggests a new way to uncover novel drug targets for sleeping sickness. In this study, we developed a novel computational method TFPP to recognize flagellar proteins in T. brucei. TFPP effectively identifies a large number of flagellar proteins with high confidence, many of which are reported first time in our study. Expression profiles of the flagellar proteome show that ,45 flagellar proteins are significantly regulated during life cycle, indicating life cycle stage-specific regulation of flagellar functions in T. brucei. We further developed a web server for TFPP with free access. Therefore, TFPP willTFPP: Trypanosome Flagellar Protein PredictorTable 2. Differential expression of the flagellar proteome in long slender (LS), short stumpy (SS) and procyclic (PC) form T. brucei.Supporting InformationTable S1 List of positive and negative samples.(DOC)Table S2 List of initial features and the element number of eachLS/SS Significantly regulated genes Up-regulatedSS/PC 289 92 197 767 (SS)LS/PC 256 125 131 760 (PC)Total93 80 13 782 (LS)feature. (DOC)Table S3 Features selected to build the final classifier model.Down-regulated3 Genes expressed(DOC)Table S4 Prediction performance of SVMaac on 50 test sets.total number of significantly regulated genes. up-regulated in 26001275 LS compared with SS, SS compared with PC, and LS compared with PC. 3 down-regulated in LS.Previous studies, the single flagellum of T. brucei changes in morphology and function during the life cycle alternating between a tsetse fly and a mammal host [3,37]. A recent study analyzed the expression profiles of the T. brucei proteome at three important life-cycle stages, namely long slender and short stumpy bloodstream forms in the mammalian host and the procyclic form in the midgut of the tsetse fly [38]. Considering uniquely mapped reads, 800 (83.4 ) flagellar genes were detected to be expressed in at least one of the three stages. Differentially expressed genes were defined to be those two-fold up- or downregulated at two stages with p0:001 according to the Audic and Claverie test [39]. In total, 363 flagellar protein-encoded genes significantly changed expression levels in at least one of the three stages, accounting for ,45 of the expressed flagellar proteins (Table 2). As expected, much more flagellar genes changed their expression levels in procyclic form when compared with the other two bloodstream stages. As T. brucei lives in from the tsetse fly to the mammal host, the parasite needs more genes to be regulated to adapt to host change for survival. We found that most of these differentially expressed genes were up-regulated in the procyclic form when compared with the long slender and short stumpybloodstream form. This is not surprising, as we know that the flagellum-mediated migration between the midgut and salivary glands of its tsetse fly vector is essential for the progression of its life cycle [3,40]. When compared with the short stumpy bloodstream form, we found much more flagellar genes were up-regulated in the long slender and procyclic forms. This may due to the important roles of the single flagellum in cell division as demonstrated by previous studies [5,40,41,42], while both the long slender bloodstream form and the procyclic form are proliferative forms. These results indicate life cycle stage-specific regulation of flagellar functions in T. brucei.ConclusionsThe available evidence indicates the multifunctional nature of the single flagellum in T. brucei, and suggests a new way to uncover novel drug targets for sleeping sickness. In this study, we developed a novel computational method TFPP to recognize flagellar proteins in T. brucei. TFPP effectively identifies a large number of flagellar proteins with high confidence, many of which are reported first time in our study. Expression profiles of the flagellar proteome show that ,45 flagellar proteins are significantly regulated during life cycle, indicating life cycle stage-specific regulation of flagellar functions in T. brucei. We further developed a web server for TFPP with free access. Therefore, TFPP willTFPP: Trypanosome Flagellar Protein PredictorTable 2. Differential expression of the flagellar proteome in long slender (LS), short stumpy (SS) and procyclic (PC) form T. brucei.Supporting InformationTable S1 List of positive and negative samples.(DOC)Table S2 List of initial features and the element number of eachLS/SS Significantly regulated genes Up-regulatedSS/PC 289 92 197 767 (SS)LS/PC 256 125 131 760 (PC)Total93 80 13 782 (LS)feature. (DOC)Table S3 Features selected to build the final classifier model.Down-regulated3 Genes expressed(DOC)Table S4 Prediction performance of SVMaac on 50 test sets.total number of significantly regulated genes. up-regulated in 26001275 LS compared with SS, SS compared with PC, and LS compared with PC. 3 down-regulated in LS.

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