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Title: The role of light in photosynthetic cyanophages : from physiology to gene expression
Author: Puxty, Richard John
ISNI:       0000 0004 5348 1637
Awarding Body: University of Warwick
Current Institution: University of Warwick
Date of Award: 2014
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It is estimated that there are approximately 1030 ocean virioplankton (Suttle 2007; Parsons et al. 2012). A large component of the oceanic viriosphere are the cyanophages, viruses that specifically infect cyanobacteria. Recent advances in genomics has revealed such viruses encode a multitude of genes, often acquired horizontally, that act to redirect metabolism for their own gains (Mann et al. 2003; Lindell et al. 2004a; Millard et al. 2009; Sullivan et al. 2010; Hurwitz et al. 2013; Enav et al. 2014). These genes have been named auxiliary metabolic genes (AMGs). They include multiple subunits of complexes involved with photosynthetic electron transport (PET) and CO2 fixation (Mann et al. 2003; Lindell et al. 2004; Millard et al. 2009; Sullivan et al. 2010; Thompson et al. 2011; Puxty et al. submitted), leading to the hypothesis that cyanophages directly participate in photosynthesis to provide carbon and energy for their own replication. Cyanophages face a dynamically changing light environment during their rather lengthy infection cycles ~12hrs. Therefore, it was hypothesised that changes in light intensity may affect the physiology of phage infection in terms of photosynthesis, CO2 fixation and infection dynamics. During infection of the marine cyanobacterium Synechococcus sp. WH7803 with the well characterised cyanophage S-PM2 I show that decoupling of the photochemical and CO2 fixation reactions of photosynthesis occurs (Chapter 3), which presumably redirects metabolism towards energy generation and away from growth. Moreover, S-PM2 acts to modify the PET which results in improved functioning of PSII at HL. The result is that the lytic cycle is significantly shortened during infection of the Synechococcus host under HL compared with low light (LL) conditions. To understand whether this early lysis is a regulated process, whole transcriptome sequencing of S-PM2 was performed in HL and LL (Chapter 5). This revealed a general increase in expression of all genes in HL but only the cyanophage psbA gene was significantly up-regulated above this background. This AMG encodes a core complex of photosystem II (PSII) of the PET and therefore plays a vital role in supplying energy through photophosphorylation. It is concluded that light poses a metabolic constraint on cyanophage development that requires large amounts of energy for synthesis and assembly of the structural components of the virion. Cyanophages have therefore acquired and evolved coordinated expression of PSII genes to maintain this supply of energy. I further hypothesise that gene expression may pose a significant barrier in the acquisition of AMGs from their host due to incompatible gene regulation. To test this, the phage transcriptome was analysed (Chapter 4) to validate the model of temporal transcriptional regulation in cyanophage S-PM2 as previously proposed by comparison to enterobacteriophage T4. It is shown that the experimental data is largely congruent with the proposed model. This also revealed unpredicted characteristics of the transcriptome, including genome wide transcriptional read-through and antisense expression. It is suggested that this is facilitated by either inefficient transcriptional termination or pervasive transcription initiation and may be a biologically relevant process that allows for moderate expression of recently acquired genes. In addition, genome-wide antisense transcription may act to regulate the inventory or temporal expression of specific mRNAs in these regulatory limited phages. Attempts were therefore made to characterise a previously detected non-coding RNA (ncRNA) antisense to the light regulated S-PM2 psbA gene (Chapter 6). A model is proposed suggesting that the asRNA may act to tweak psbA expression under LL conditions to prevent accumulation of unnecessary PSII proteins. This mechanism has an interesting effect on the rate of splicing of a group I intron encoded by the psbA gene. This study provides an important leap forward in our understanding of the factors that regulate the infection dynamics and therefore ecology of cyanophages. In so doing it also reveals transcriptional constraints and adaptations that go some way to explaining the evolution of cyanophage genomes.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QH Natural history ; QR355 Virology