A Quantitative Proteomic Analysis of Light Adaptation in a Globally Significant Marine Cyanobacterium <i>Prochlorococcus marinus</i> MED4

Pandhal, Jagroop; Wright, Phillip C.; Biggs, Catherine A.

doi:10.1021/pr060460c

Cited by 37 publications

(49 citation statements)

References 58 publications

(153 reference statements)

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“…The downregulation of photosystem-and respiration-related proteins under conditions of Fe limitation reduced the overall growth of the cultures compared to that of their Fe-replete counterparts. These general housekeeping responses have been well documented in other cyanobacterial studies in which the effects of light stress and nutrient limitation were examined (64,65).…”

Section: Discussionmentioning

confidence: 62%

Physiological and Proteomic Responses of Continuous Cultures of Microcystis aeruginosa PCC 7806 to Changes in Iron Bioavailability and Growth Rate

Yeung

D’Agostino

Poljak

et al. 2016

Appl Environ Microbiol

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The hepatotoxin microcystin (MCYST) is produced by a variety of freshwater cyanobacterial species, including Microcystis aeruginosa. Interestingly, MCYST-producing M. aeruginosa strains have been shown to outcompete their nontoxic counterparts under iron-limiting conditions. However, the reasons for this are unclear. Here we examined the proteomic response of M. aeruginosa PCC 7806 continuous cultures under different iron and growth regimes. Iron limitation was correlated with a global reduction in levels of proteins associated with energy metabolism and photosynthesis. These proteomic changes were consistent with physiological observations, including reduced chlorophyll a content and reduced cell size. While levels of MCYST biosynthesis proteins did not fluctuate during the study period, both intra-and extracellular toxin quotas were significantly higher under iron-limiting conditions. Our results support the hypothesis that intracellular MCYST plays a role in protecting the cell against oxidative stress. Further, we propose that extracellular MCYST may act as a signaling molecule, stimulating MCYST production under conditions of iron limitation and enhancing the fitness of bloom populations. IMPORTANCEMicrocystin production in water supply reservoirs is a global public health problem. Understanding the ecophysiology of hepatotoxic cyanobacteria, including their responses to the presence of key micronutrient metals such as iron, is central to managing harmful blooms. To our knowledge, this was the first study to examine proteomic and physiological changes occurring in M. aeruginosa continuous cultures under conditions of iron limitation at different growth rates. C yanobacteria ("blue-green algae") proliferate in warm stratified water bodies rich in nitrogen and phosphorus (1, 2). Cyanobacterial blooms can have a negative impact on the appearance, taste, and odor of water, and their subsequent decay can lead to oxygen depletion and fish kills (3). However, of greatest concern to public health is their production of potent toxins. Microcystis aeruginosa (Kützing) Lemmermann is a common bloom-forming cyanobacterial species that produces hepatotoxic microcystins (MCYSTs) (1). These nonribosomally synthesized peptides inhibit eukaryotic protein phosphatases, leading to liver necrosis in acute doses and hepatocellular carcinoma in chronic low doses (4, 5).There is strong evidence that the production of MCYST is a direct function of cell division (6). It is therefore not surprising that parameters affecting cyanobacterial growth rate (e.g., nutrients, trace metals, temperature, pH, and light) have been correlated with fluctuating MCYST levels in batch culture experiments (7-9).Recent molecular studies suggest that iron (Fe) and nitrogen (N) also play a role in regulating the expression of MCYST biosynthesis (mcy) genes. In M. aeruginosa, for example, the mcy promoter region contains binding sites for the ferric uptake regulator (Fur) and the global nitrogen regulator (NtcA) (10, 11). Despite these genetic clu...

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Section: Discussionmentioning

confidence: 62%

Physiological and Proteomic Responses of Continuous Cultures of Microcystis aeruginosa PCC 7806 to Changes in Iron Bioavailability and Growth Rate

Yeung

D’Agostino

Poljak

et al. 2016

Appl Environ Microbiol

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“…The quantitative comparison of differentially regulated proteins revealed a significantly reduced expression of photosystem-related genes upon high-light intensities, which was attributed to the prevention of light-induced damage. By contrast, the general stress proteins GroEL, HtpG and GroES were strongly up-regulated under high-light conditions, implicating that a protective response to stress caused by high irradiances is required [43].…”

Section: Cyanobacteriamentioning

confidence: 91%

“…Varying levels of light intensity iTRAQ and LC-M/MS [43] Heterotrophy/photoautotrophy 2-D PAGE, MALDI-MS and LC-MS/MS [50] Nutrient availability/different C-and energy sources [68,74,75] Growth phase/cell cycle…”

Section: Proteome Signatures In Response To Changing Environmental Comentioning

confidence: 99%

“…As the genomes of five individual Prochlorococcus strains have been fully sequenced, genomic as well as proteomic analyses of their peculiar light-adaptation strategies have become feasible. In a recent study, cultures of the high-light-adapted P. marinus strain MED4 were exposed to varying levels of light intensity and the resulting protein patterns were compared using the isobaric tags for relative and absolute quantification (iTRAQ) method [43]. In total, 184 Prochlorococcus proteins could be identified and characterized as to their function, pI, M r and cellular location.…”

Section: Cyanobacteriamentioning

confidence: 99%

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Proteomics of marine bacteria

2008

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Little is known about the life of marine microorganisms under their particular environmental conditions. Genome sequencing combined with the techniques of functional genomics, especially proteome analyses, now open up revolutionary insights into the adaptation strategies marine organisms have evolved in response to the challenges of their habitat. This report summarizes the first approaches and state‐of‐the‐art in the field of proteome analysis of marine bacteria. This includes, amongst others, proteomics on culturable, free‐living marine bacteria and on uncultivable bacteria living in symbiosis with higher organisms. Finally, new approaches to determine the metaproteome of uncultured microbial consortia from marine habitats are discussed.

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“…Peptide fractionation was achieved using strong cation exchange (SCX) on a BioLC HPLC (Dionex, Surrey, UK). The gradients, buffers, and conditions have been described previously (Pandhal et al, 2007). analysis of system level changes is followed by a data-driven ''design stage'' from discovery analysis.…”

Section: Itraq Labeling and Peptide Fractionationmentioning

confidence: 99%

Improving N‐glycosylation efficiency in Escherichia coli using shotgun proteomics, metabolic network analysis, and selective reaction monitoring

Pandhal

Noirel

et al. 2010

Biotech & Bioengineering

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Recently, the prospect of using Escherichia coli as a host for human glycoprotein production has increased due to detailed characterization of the prokaryotic N-glycosylation process and the ability to transfer the system into this bacterium. Although functionality of the native Campylobacter jejuni N-glycosylation system in E. coli has been demonstrated, the efficiency of the process using the well-characterized C. jejuni glycoprotein AcrA, was found to be low at 13.4±0.9% of total extracted protein. A combined approach using isobaric labeling of peptides and probability-based network analysis of metabolic changes was applied to forward engineer E. coli to improve glycosylation efficiency of AcrA. Enhancing flux through the glyoxylate cycle was identified as a potential metabolic manipulation to improve modification efficiency and was achieved by increasing the expression of isocitrate lyase. While the overall recombinant protein titre did not change significantly, the amount of glycosylated protein increased by approximately 300%.

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A Quantitative Proteomic Analysis of Light Adaptation in a Globally Significant Marine Cyanobacterium Prochlorococcus marinus MED4

Cited by 37 publications

References 58 publications

Physiological and Proteomic Responses of Continuous Cultures of Microcystis aeruginosa PCC 7806 to Changes in Iron Bioavailability and Growth Rate

Physiological and Proteomic Responses of Continuous Cultures of Microcystis aeruginosa PCC 7806 to Changes in Iron Bioavailability and Growth Rate

Proteomics of marine bacteria

Improving N‐glycosylation efficiency in Escherichia coli using shotgun proteomics, metabolic network analysis, and selective reaction monitoring

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