Project Details
Projekt
The role of m6A-RNA modification to control phage replication
Applicants
Dr. Sophie Brameyer; Professorin Dr. Kirsten Jung
Subject Area
Metabolism, Biochemistry and Genetics of Microorganisms
Term
since 2021
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 464582101
In eukaryotes, viral replication is controlled by the methylation of messenger RNA (mRNA). In particular, newly transcribed viral RNAs undergo post-transcriptional N6-adenosine (m6A) methylation. In addition, viral infection leads to an increase of a m6A modification of the host mRNA, which limits viral replication. So far, almost nothing is known about the role of mRNA modification in controlling phage replication in prokaryotes. In this project we will elucidate the role of m6A-RNA methylation in phage replication in the marine bacterium Vibrio campbellii ATCC BAA-116 (formerly V. harveyi BB120), a model organism for quorum sensing. In our preliminary work we have been able to show that V. campbellii contains four prophages, and oxidative stress initiates the lytic cycle. As an important prerequisite for this project we isolated a m6A-RNA-depleted V. campbellii mutant in which two genes (rlmF and rlmJ) coding for methyltransferases are deleted. This mutant is characterized by a significantly higher degree of prophage induction and cell lysis compared to the wild type suggesting that also in prokaryotes RNA methylation plays a role in controlling phage replication.In the first work package we will compare the wild type and a m6A-RNA depleted mutant (∆rlmF∆rlmJ) with respect to the induction of the lytic cycle of the four prophages in V. campbellii under different conditions (UV stress, heat, cell density, microbiota and microalgae derived molecules). Furthermore, the influence of m6A-RNA methylation on the infection potential (lysis or lysogeny) of available and newly isolated phages will be tested. In the second work package we will investigate the influence of m6A-RNA methylation on the decision of single cells to activate prophages or to resist lytic phages. We will analyze fluorescent transcriptional and translational reporter fusions of the two intact prophages encoded in the two V. campbellii chromosomes and spatio-temporal changes in the life cycle of the phages at single cell level. In the third work package we will investigate to what extent phage replication influences the m6A methylation pattern in V. campbellii.This project addresses a previously unexplored regulatory level, the translation level, in the replication cycle of phages. We expect to gain new insights into phage-bacteria interaction in general and specifically into the fine-tuning of the decision making process of individual cells. Our long-term goal is to search for effectors that modulate m6A-RNA modification to identify novel antiviral compounds.
DFG Programme
Priority Programmes
