Poster abstracts
Poster number 27 submitted by Carli Camporeale
Nucleotide-resolution Mapping of RNA N6-Methyladenosine (m6A) in Malaria Pathogen Plasmodium falciparum and Determining m6A’s Role in Host–Pathogen Interactions
Carli Camporeale (Department of Biological Sciences, University of North Carolina, Charlotte, NC, USA, United States of America), Cassie Catacalos, Kaitlin Klotz (Department of Biological Sciences, University of North Carolina, Charlotte, NC, USA, United States of America), Doaa Hasan, Sahiti Shomalraju, Sarath Chandra Janga (Department of BioHealth Informatics, Luddy School of Informatics, Computing and Engineering, Indiana University Indianapolis IUI, Indianapolis, Indiana, United States of America), Matthew Tegowski, Kate Meyer (Department of Biochemistry, Duke University School of Medicine, Durham, North Carolina, United States of America), Arthur Hunt (Department of Plant and Soil Sciences, University of Kentucky, Lexington, KY, USA, United States of America), Kausik Chakrabarti (Department of Biological Sciences, University of North Carolina, Charlotte, NC, USA, United States of America)
Abstract:
The expression of a precise mRNA transcriptome is crucial for establishing cell function. To this end, the N6-methyladenosine (m6A) modification is emerging as the most prevalent and abundant chemical modification in eukaryotic mRNAs, which plays important roles in various biological processes and can regulate all aspects of mRNA metabolism, ranging from 5′ end-capping, polyadenylation, pre-mRNA splicing and translation. Thus, this chemical modification is of profound importance to eukaryotic gene expression regulation. Malaria, caused by apicomplexan parasite Plasmodium falciparum, is the deadliest vector-borne disease in the world, claiming more than 600,000 lives each year. m6A modifications have been implicated in malaria gene regulation. However, m6A’s mechanistic role in mRNA metabolism in this clinically important protist remains unknown. The survival of P. falciparum in the human host is often threatened by two important stressors – febrile temperature resulting from human innate immune response and antimalarial drugs, which induce oxidative stress. These stress conditions significantly decrease the sensitivity of the parasite to the drug as a survival response, but the underlying molecular mechanisms remain poorly understood. Since m6A’s role as a newly emerging layer of genetic control in stress responses is becoming more and more evident, here we seek to determine the extent of m6A modifications in the malaria epitranscriptome and investigate the heat and drug-induced stress response on m6A dynamicity in the malaria blood stage development in the human host. To globally map m6A modifications in P. falciparum, we used DART-seq technology to directly determine m6A sites at a single-nucleotide resolution. Additionally, we have developed an m6A mapping and detection pipeline using Oxford Nanopore-based direct RNA sequencing. To gain insight into m6A’s functional significance, we investigated m6A epitranscriptomic changes in response to a variety of stresses. These studies should shed light on the extent and dynamicity of this intrinsic RNA modification in malaria and determine m6A’s potential role in stress responses in the human host.
Keywords: m6A, RNA modifications