University of Kentucky
Alternative Polyadenylation in Sarcocystis neurona
Institution
University of Kentucky
Faculty Advisor/ Mentor
Arthur Hunt; Daniel Howe
Abstract
This project studied the occurrence of alternative polyadenylation during the growth of Sarcocystis neurona. S. neurona is an obligate intracellular parasite that causes equine protozoal myeloencephalitis. Among the stages of growth of the parasite are the merozoite and schizont stages. The merozoite is the extracellular parasite that has been lysed from the host cell once the development has completed. The schizont is the intracellular development stage, where it grows and acquires nutrients, eventually producing 64 haploid merozoites. While some gene expression analysis has been conducted, there is no knowledge on polyadenylation in S. neurona. Moreover, there have been no genome-wide studies of poly(A) site choice for any member of the Apicomplexans phylum. Thus, it is not clear if alternative polyadenylation contributes to changes in gene expression in these organisms, and any findings will add to current knowledge. This study addressed the hypothesis that, indeed, alternative polyadenylation does contribute to regulated gene expression in S. neurona. Poly(A) tags (1) were made to conduct a genome-wide study of poly(A) site choice. The tag libraries were sequenced on a MiSeq instrument, returning 11.9 million reads. The data was analyzed using programs including CLC Genomics Workbench and Bedtools, as well as others designed specifically for APA analysis to define sites and assess alternative polyadenylation. The results suggested that S. neurona possesses a distinctive polyadenylation signal that is reminiscent of that seen in higher plants. The results also showed multiple instances of APA. The results from this study confirm our hypothesis that there will be changes in poly(A) site choice during the developmental stages of S. neurona.
Alternative Polyadenylation in Sarcocystis neurona
This project studied the occurrence of alternative polyadenylation during the growth of Sarcocystis neurona. S. neurona is an obligate intracellular parasite that causes equine protozoal myeloencephalitis. Among the stages of growth of the parasite are the merozoite and schizont stages. The merozoite is the extracellular parasite that has been lysed from the host cell once the development has completed. The schizont is the intracellular development stage, where it grows and acquires nutrients, eventually producing 64 haploid merozoites. While some gene expression analysis has been conducted, there is no knowledge on polyadenylation in S. neurona. Moreover, there have been no genome-wide studies of poly(A) site choice for any member of the Apicomplexans phylum. Thus, it is not clear if alternative polyadenylation contributes to changes in gene expression in these organisms, and any findings will add to current knowledge. This study addressed the hypothesis that, indeed, alternative polyadenylation does contribute to regulated gene expression in S. neurona. Poly(A) tags (1) were made to conduct a genome-wide study of poly(A) site choice. The tag libraries were sequenced on a MiSeq instrument, returning 11.9 million reads. The data was analyzed using programs including CLC Genomics Workbench and Bedtools, as well as others designed specifically for APA analysis to define sites and assess alternative polyadenylation. The results suggested that S. neurona possesses a distinctive polyadenylation signal that is reminiscent of that seen in higher plants. The results also showed multiple instances of APA. The results from this study confirm our hypothesis that there will be changes in poly(A) site choice during the developmental stages of S. neurona.