Publications by year
2023
Brownlee C, Helliwell KE, Meeda Y, McLachlan D, Murphy EA, Wheeler GL (2023). Regulation and integration of membrane transport in marine diatoms.
Seminars in Cell and Developmental Biology,
134, 79-89.
Abstract:
Regulation and integration of membrane transport in marine diatoms
Diatoms represent one of the most successful groups of marine phytoplankton and are major contributors to ocean biogeochemical cycling. They have colonized marine, freshwater and ice environments and inhabit all regions of the World's oceans, from poles to tropics. Their success is underpinned by a remarkable ability to regulate their growth and metabolism during nutrient limitation and to respond rapidly when nutrients are available. This requires precise regulation of membrane transport and nutrient acquisition mechanisms, integration of nutrient sensing mechanisms and coordination of different transport pathways. This review outlines transport mechanisms involved in acquisition of key nutrients (N, C, P, Si, Fe) by marine diatoms, illustrating their complexity, sophistication and multiple levels of control.
Abstract.
2022
Kleiner FH, Helliwell KE, Chrachri A, Hopes A, Parry-Wilson H, Gaikwad T, Mieszkowska N, Mock T, Wheeler GL, Brownlee C, et al (2022). Cold-induced [Ca2+]cyt elevations function to support osmoregulation in marine diatoms. Plant Physiology, 190(2), 1384-1399.
Helliwell KE (2022). Emerging trends in nitrogen and phosphorus signalling in photosynthetic eukaryotes. Trends in Plant Science, 28(3), 344-358.
Kottmeier DM, Chrachri A, Langer G, Helliwell KE, Wheeler GL, Brownlee C (2022). Reduced H+ channel activity disrupts pH homeostasis and calcification in coccolithophores at low ocean pH.
Proc Natl Acad Sci U S A,
119(19).
Abstract:
Reduced H+ channel activity disrupts pH homeostasis and calcification in coccolithophores at low ocean pH.
Coccolithophores are major producers of ocean biogenic calcite, but this process is predicted to be negatively affected by future ocean acidification scenarios. Since coccolithophores calcify intracellularly, the mechanisms through which changes in seawater carbonate chemistry affect calcification remain unclear. Here we show that voltage-gated H+ channels in the plasma membrane of Coccolithus braarudii serve to regulate pH and maintain calcification under normal conditions but have greatly reduced activity in cells acclimated to low pH. This disrupts intracellular pH homeostasis and impairs the ability of C. braarudii to remove H+ generated by the calcification process, leading to specific coccolith malformations. These coccolith malformations can be reproduced by pharmacological inhibition of H+ channels. Heavily calcified coccolithophore species such as C. braarudii, which make the major contribution to carbonate export to the deep ocean, have a large intracellular H+ load and are likely to be most vulnerable to future decreases in ocean pH.
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Author URL.
Helliwell KE, Shibl AA, Amin SA (2022). The Diatom Microbiome: New Perspectives for Diatom-Bacteria Symbioses. In (Ed) The Molecular Life of Diatoms, 679-712.
2021
Helliwell KE, Harrison EL, Christie-Oleza JA, Rees AP, Kleiner FH, Gaikwad T, Downe J, Aguilo-Ferretjans MM, Al-Moosawi L, Brownlee C, et al (2021). A Novel Ca2+ Signaling Pathway Coordinates Environmental Phosphorus Sensing and Nitrogen Metabolism in Marine Diatoms. Current Biology, 31(5), 978-989.e4.
Helliwell KE, Kleiner FH, Hardstaff H, Chrachri A, Gaikwad T, Salmon D, Smirnoff N, Wheeler GL, Brownlee C (2021). Spatiotemporal patterns of intracellular Ca<sup>2+</sup> signalling govern hypo‐osmotic stress resilience in marine diatoms.
New Phytologist,
230(1), 155-170.
Abstract:
Spatiotemporal patterns of intracellular Ca2+ signalling govern hypo‐osmotic stress resilience in marine diatoms
Summary
Diatoms are globally important phytoplankton that dominate coastal and polar‐ice assemblages. These environments exhibit substantial changes in salinity over dynamic spatiotemporal regimes. Rapid sensory systems are vital to mitigate the harmful consequences of osmotic stress. Population‐based analyses have suggested that Ca2+ signalling is involved in diatom osmotic sensing. However, mechanistic insight of the role of osmotic Ca2+ signalling is limited.
Here, we show that Phaeodactylum Ca2+ elevations are essential for surviving hypo‐osmotic shock. Moreover, employing novel single‐cell imaging techniques we have characterised real‐time Ca2+ signalling responses in single diatom cells to environmental osmotic perturbations.
We observe that intracellular spatiotemporal patterns of osmotic‐induced Ca2+ elevations encode vital information regarding the nature of the osmotic stimulus. Localised Ca2+ signals evoked by mild or gradual hypo‐osmotic shocks are propagated globally from the apical cell tips, enabling fine‐tuned cell volume regulation across the whole cell.
Finally, we demonstrate that diatoms adopt Ca2+‐independent and dependent mechanisms for osmoregulation. We find that efflux of organic osmolytes occurs in a Ca2+‐independent manner, but this response is insufficient to mitigate cell damage during hypo‐osmotic shock. By comparison, Ca2+‐dependent signalling is necessary to prevent cell bursting via precise coordination of K+ transport, and therefore is likely to underpin survival in dynamic osmotic environments.
Abstract.
2020
Helliwell KE, Chrachri A, Koester JA, Wharam S, Taylor AR, Wheeler GL, Brownlee C (2020). A Novel Single-Domain Na+-Selective Voltage-Gated Channel in Photosynthetic Eukaryotes.
Plant Physiol,
184(4), 1674-1683.
Abstract:
A Novel Single-Domain Na+-Selective Voltage-Gated Channel in Photosynthetic Eukaryotes.
The evolution of Na+-selective four-domain voltage-gated channels (4D-Navs) in animals allowed rapid Na+-dependent electrical excitability, and enabled the development of sophisticated systems for rapid and long-range signaling. While bacteria encode single-domain Na+-selective voltage-gated channels (BacNav), they typically exhibit much slower kinetics than 4D-Navs, and are not thought to have crossed the prokaryote-eukaryote boundary. As such, the capacity for rapid Na+-selective signaling is considered to be confined to certain animal taxa, and absent from photosynthetic eukaryotes. Certainly, in land plants, such as the Venus flytrap (Dionaea muscipula) where fast electrical excitability has been described, this is most likely based on fast anion channels. Here, we report a unique class of eukaryotic Na+-selective, single-domain channels (EukCatBs) that are present primarily in haptophyte algae, including the ecologically important calcifying coccolithophores, Emiliania huxleyi and Scyphosphaera apsteinii the EukCatB channels exhibit very rapid voltage-dependent activation and inactivation kinetics, and isoform-specific sensitivity to the highly selective 4D-Nav blocker tetrodotoxin. The results demonstrate that the capacity for rapid Na+-based signaling in eukaryotes is not restricted to animals or to the presence of 4D-Navs. The EukCatB channels therefore represent an independent evolution of fast Na+-based electrical signaling in eukaryotes that likely contribute to sophisticated cellular control mechanisms operating on very short time scales in unicellular algae.
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Author URL.
Bunbury F, Helliwell KE, Mehrshahi P, Davey MP, Salmon DL, Holzer A, Smirnoff N, Smith AG (2020). Responses of a Newly Evolved Auxotroph of Chlamydomonas to B12 Deprivation.
Plant Physiol,
183(1), 167-178.
Abstract:
Responses of a Newly Evolved Auxotroph of Chlamydomonas to B12 Deprivation.
The corrinoid B12 is synthesized only by prokaryotes yet is widely required by eukaryotes as an enzyme cofactor. Microalgae have evolved B12 dependence on multiple occasions, and we previously demonstrated that experimental evolution of the non-B12-requiring alga Chlamydomonas reinhardtii in media supplemented with B12 generated a B12-dependent mutant (hereafter metE7). This clone provides a unique opportunity to study the physiology of a nascent B12 auxotroph. Our analyses demonstrate that B12 deprivation of metE7 disrupts C1 metabolism, causes an accumulation of starch and triacylglycerides, and leads to a decrease in photosynthetic pigments, proteins, and free amino acids. B12 deprivation also caused a substantial increase in reactive oxygen species, which preceded rapid cell death. Survival could be improved without compromising growth by simultaneously depriving the cells of nitrogen, suggesting a type of cross protection. Significantly, we found further improvements in survival under B12 limitation and an increase in B12 use efficiency after metE7 underwent a further period of experimental evolution, this time in coculture with a B12-producing bacterium. Therefore, although an early B12-dependent alga would likely be poorly adapted to coping with B12 deprivation, association with B12-producers can ensure long-term survival whilst also providing a suitable environment for evolving mechanisms to tolerate B12 limitation better.
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Author URL.
2019
Chrachri A, Koester J, Wharam S, Verret F, Taylor A, Wheeler G, Brownlee C, Helliwell K (2019). Alternative mechanisms for fast Na+/Ca2+ signalling in eukaryotes via a novel class of single-domain voltage-gated channels. Current Biology, 29, 1505-1505.
Wheeler G, Helliwell K, Brownlee C (2019). Calcium signalling in algae. Perspectives in Phycology, 6(1-2), 1-10.
Leebens-Mack JH, Barker MS, Carpenter EJ, Deyholos MK, Gitzendanner MA, Graham SW, Grosse I, Li Z, Melkonian M, Mirarab S, et al (2019). One thousand plant transcriptomes and the phylogenomics of green plants.
Nature,
574(7780), 679-685.
Abstract:
One thousand plant transcriptomes and the phylogenomics of green plants
Green plants (Viridiplantae) include around 450,000–500,000 species1,2 of great diversity and have important roles in terrestrial and aquatic ecosystems. Here, as part of the One Thousand Plant Transcriptomes Initiative, we sequenced the vegetative transcriptomes of 1,124 species that span the diversity of plants in a broad sense (Archaeplastida), including green plants (Viridiplantae), glaucophytes (Glaucophyta) and red algae (Rhodophyta). Our analysis provides a robust phylogenomic framework for examining the evolution of green plants. Most inferred species relationships are well supported across multiple species tree and supermatrix analyses, but discordance among plastid and nuclear gene trees at a few important nodes highlights the complexity of plant genome evolution, including polyploidy, periods of rapid speciation, and extinction. Incomplete sorting of ancestral variation, polyploidization and massive expansions of gene families punctuate the evolutionary history of green plants. Notably, we find that large expansions of gene families preceded the origins of green plants, land plants and vascular plants, whereas whole-genome duplications are inferred to have occurred repeatedly throughout the evolution of flowering plants and ferns. The increasing availability of high-quality plant genome sequences and advances in functional genomics are enabling research on genome evolution across the green tree of life.
Abstract.
2018
Cooper M, Kazamia E, Helliwell K, Kudahl J, Sayer A, Wheeler G, Smith A (2018). Cross-exchange of B-vitamins underpins a mutualistic interaction between Ostreococcus tauri and Dinoroseobacter shibae. ISME Journal, 13, 334-345.
Helliwell KE, Pandhal J, Cooper MB, Longworth J, Kudahl UJ, Russo DA, Tomsett EV, Bunbury F, Salmon DL, Smirnoff N, et al (2018). Quantitative proteomics of a B12 -dependent alga grown in coculture with bacteria reveals metabolic tradeoffs required for mutualism.
New Phytol,
217(2), 599-612.
Abstract:
Quantitative proteomics of a B12 -dependent alga grown in coculture with bacteria reveals metabolic tradeoffs required for mutualism.
The unicellular green alga Lobomonas rostrata requires an external supply of vitamin B12 (cobalamin) for growth, which it can obtain in stable laboratory cultures from the soil bacterium Mesorhizobium loti in exchange for photosynthate. We investigated changes in protein expression in the alga that allow it to engage in this mutualism. We used quantitative isobaric tagging (iTRAQ) proteomics to determine the L. rostrata proteome grown axenically with B12 supplementation or in coculture with M. loti. Data are available via ProteomeXchange (PXD005046). Using the related Chlamydomonas reinhardtii as a reference genome, 588 algal proteins could be identified. Enzymes of amino acid biosynthesis were higher in coculture than in axenic culture, and this was reflected in increased amounts of total cellular protein and several free amino acids. A number of heat shock proteins were also elevated. Conversely, photosynthetic proteins and those of chloroplast protein synthesis were significantly lower in L. rostrata cells in coculture. These observations were confirmed by measurement of electron transfer rates in cells grown under the two conditions. The results indicate that, despite the stability of the mutualism, L. rostrata experiences stress in coculture with M. loti, and must adjust its metabolism accordingly.
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Author URL.
2017
Wells ML, Potin P, Craigie JS, Raven JA, Merchant SS, Helliwell KE, Smith AG, Camire ME, Brawley SH (2017). Algae as nutritional and functional food sources: revisiting our understanding.
Journal of Applied Phycology,
29(2), 949-982.
Abstract:
Algae as nutritional and functional food sources: revisiting our understanding
Global demand for macroalgal and microalgal foods is growing, and algae are increasingly being consumed for functional benefits beyond the traditional considerations of nutrition and health. There is substantial evidence for the health benefits of algal-derived food products, but there remain considerable challenges in quantifying these benefits, as well as possible adverse effects. First, there is a limited understanding of nutritional composition across algal species, geographical regions, and seasons, all of which can substantially affect their dietary value. The second issue is quantifying which fractions of algal foods are bioavailable to humans, and which factors influence how food constituents are released, ranging from food preparation through genetic differentiation in the gut microbiome. Third is understanding how algal nutritional and functional constituents interact in human metabolism. Superimposed considerations are the effects of harvesting, storage, and food processing techniques that can dramatically influence the potential nutritive value of algal-derived foods. We highlight this rapidly advancing area of algal science with a particular focus on the key research required to assess better the health benefits of an alga or algal product. There are rich opportunities for phycologists in this emerging field, requiring exciting new experimental and collaborative approaches.
Abstract.
Brawley SH, Blouin NA, Ficko-Blean E, Wheeler GL, Lohr M, Goodson HV, Jenkins JW, Blaby-Haas CE, Helliwell KE, Chan CX, et al (2017). Insights into the red algae and eukaryotic evolution from the genome of Porphyra umbilicalis (Bangiophyceae, Rhodophyta).
Proc Natl Acad Sci U S A,
114(31), E6361-E6370.
Abstract:
Insights into the red algae and eukaryotic evolution from the genome of Porphyra umbilicalis (Bangiophyceae, Rhodophyta).
Porphyra umbilicalis (laver) belongs to an ancient group of red algae (Bangiophyceae), is harvested for human food, and thrives in the harsh conditions of the upper intertidal zone. Here we present the 87.7-Mbp haploid Porphyra genome (65.8% G + C content, 13,125 gene loci) and elucidate traits that inform our understanding of the biology of red algae as one of the few multicellular eukaryotic lineages. Novel features of the Porphyra genome shared by other red algae relate to the cytoskeleton, calcium signaling, the cell cycle, and stress-tolerance mechanisms including photoprotection. Cytoskeletal motor proteins in Porphyra are restricted to a small set of kinesins that appear to be the only universal cytoskeletal motors within the red algae. Dynein motors are absent, and most red algae, including Porphyra, lack myosin. This surprisingly minimal cytoskeleton offers a potential explanation for why red algal cells and multicellular structures are more limited in size than in most multicellular lineages. Additional discoveries further relating to the stress tolerance of bangiophytes include ancestral enzymes for sulfation of the hydrophilic galactan-rich cell wall, evidence for mannan synthesis that originated before the divergence of green and red algae, and a high capacity for nutrient uptake. Our analyses provide a comprehensive understanding of the red algae, which are both commercially important and have played a major role in the evolution of other algal groups through secondary endosymbioses.
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Author URL.
Helliwell KE (2017). Tansley insight: the roles of B vitamins in phytoplankton nutrition: new perspectives and prospects.
New Phytologist,
216(1), 62-68.
Abstract:
Tansley insight: the roles of B vitamins in phytoplankton nutrition: new perspectives and prospects
B vitamins play essential roles in central metabolism. These organic water-soluble molecules act as, or as part of, coenzymes within the cell. Unlike land plants, many eukaryotic algae are auxotrophic for certain B vitamins. Recent progress in algal genetic resources and environmental chemistry have promoted a renewal of interest in the role of vitamins in governing phytoplankton dynamics, and illuminated amazing versatility in phytoplankton vitamin metabolism. Accumulating evidence demonstrates metabolic complexity in the production and bioavailability of different vitamin forms, coupled with specialized acquisition strategies to salvage and remodel vitamin precursors. Here, I describe recent advances and discuss how they redefine our view of the way in which vitamins are cycled in aquatic ecosystems and their importance in structuring phytoplankton communities.
Abstract.
2016
Helliwell KE, Lawrence AD, Holzer A, Kudahl UJ, Sasso S, Kräutler B, Scanlan DJ, Warren MJ, Smith AG (2016). Cyanobacteria and Eukaryotic Algae Use Different Chemical Variants of Vitamin B<inf>12</inf>.
Current Biology,
26(8), 999-1008.
Abstract:
Cyanobacteria and Eukaryotic Algae Use Different Chemical Variants of Vitamin B12
Eukaryotic microalgae and prokaryotic cyanobacteria are the major components of the phytoplankton. Determining factors that govern growth of these primary producers, and how they interact, is therefore essential to understanding aquatic ecosystem productivity. Over half of microalgal species representing marine and freshwater habitats require for growth the corrinoid cofactor B12, which is synthesized de novo only by certain prokaryotes, including the majority of cyanobacteria. There are several chemical variants of B12, which are not necessarily functionally interchangeable. Cobalamin, the form bioavailable to humans, has as its lower axial ligand 5,6-dimethylbenzimidazole (DMB). Here, we show that the abundant marine cyanobacterium Synechococcus synthesizes only pseudocobalamin, in which the lower axial ligand is adenine. Moreover, bioinformatic searches of over 100 sequenced cyanobacterial genomes for B12 biosynthesis genes, including those involved in nucleotide loop assembly, suggest this is the form synthesized by cyanobacteria more broadly. We further demonstrate that pseudocobalamin is several orders of magnitude less bioavailable than cobalamin to several B12-dependent microalgae representing diverse lineages. This indicates that the two major phytoplankton groups use a different B12 currency. However, in an intriguing twist, some microalgal species can use pseudocobalamin if DMB is provided, suggesting that they are able to remodel the cofactor, whereas Synechococcus cannot. This species-specific attribute implicates algal remodelers as novel and keystone players of the B12 cycle, transforming our perception of the dynamics and complexity of the flux of this nutrient in aquatic ecosystems.
Abstract.
Kazamia E, Helliwell KE, Purton S, Smith AG (2016). How mutualisms arise in phytoplankton communities: building eco-evolutionary principles for aquatic microbes.
Ecology letters,
19(7), 810-822.
Abstract:
How mutualisms arise in phytoplankton communities: building eco-evolutionary principles for aquatic microbes
Extensive sampling and metagenomics analyses of plankton communities across all aquatic environments are beginning to provide insights into the ecology of microbial communities. In particular, the importance of metabolic exchanges that provide a foundation for ecological interactions between microorganisms has emerged as a key factor in forging such communities. Here we show how both studies of environmental samples and physiological experimentation in the laboratory with defined microbial co-cultures are being used to decipher the metabolic and molecular underpinnings of such exchanges. In addition, we explain how metabolic modelling may be used to conduct investigations in reverse, deducing novel molecular exchanges from analysis of large-scale data sets, which can identify persistently co-occurring species. Finally, we consider how knowledge of microbial community ecology can be built into evolutionary theories tailored to these species' unique lifestyles. We propose a novel model for the evolution of metabolic auxotrophy in microorganisms that arises as a result of symbiosis, termed the Foraging-to-Farming hypothesis. The model has testable predictions, fits several known examples of mutualism in the aquatic world, and sheds light on how interactions, which cement dependencies within communities of microorganisms, might be initiated.
Abstract.
Nguyen GTDT, Scaife MA, Helliwell KE, Smith AG (2016). Role of riboswitches in gene regulation and their potential for algal biotechnology.
Journal of Phycology,
52(3), 320-328.
Abstract:
Role of riboswitches in gene regulation and their potential for algal biotechnology
Riboswitches are regulatory elements in messenger RNA to which specific ligands can bind directly in the absence of proteins. Ligand binding alters the mRNA secondary structure, thereby affecting expression of the encoded protein. Riboswitches are widespread in prokaryotes, with over 20 different effector ligands known, including amino acids, cofactors, and Mg2+ ions, and gene expression is generally regulated by affecting translation or termination of transcription. In plants, fungi, and microalgae, riboswitches have been found, but only those that bind thiamine pyrophosphate. These eukaryotic riboswitches operate by causing alternative splicing of the transcript. Here, we review the current status of riboswitch research with specific emphasis on microalgae. We discuss new riboswitch discoveries and insights into the underlying mechanism of action, and how next generation sequencing technology provides the motivation and opportunity to improve our understanding of these rare but important regulatory elements. We also highlight the potential of microalgal riboswitches as a tool for synthetic biology and industrial biotechnology.
Abstract.
2015
Scaife MA, Nguyen GTDT, Rico J, Lambert D, Helliwell KE, Smith AG (2015). Establishing Chlamydomonas reinhardtii as an industrial biotechnology host.
Plant Journal,
82(3), 532-546.
Abstract:
Establishing Chlamydomonas reinhardtii as an industrial biotechnology host
Summary Microalgae constitute a diverse group of eukaryotic unicellular organisms that are of interest for pure and applied research. Owing to their natural synthesis of value-added natural products microalgae are emerging as a source of sustainable chemical compounds, proteins and metabolites, including but not limited to those that could replace compounds currently made from fossil fuels. For the model microalga, Chlamydomonas reinhardtii, this has prompted a period of rapid development so that this organism is poised for exploitation as an industrial biotechnology platform. The question now is how best to achieve this? Highly advanced industrial biotechnology systems using bacteria and yeasts were established in a classical metabolic engineering manner over several decades. However, the advent of advanced molecular tools and the rise of synthetic biology provide an opportunity to expedite the development of C. reinhardtii as an industrial biotechnology platform, avoiding the process of incremental improvement. In this review we describe the current status of genetic manipulation of C. reinhardtii for metabolic engineering. We then introduce several concepts that underpin synthetic biology, and show how generic parts are identified and used in a standard manner to achieve predictable outputs. Based on this we suggest that the development of C. reinhardtii as an industrial biotechnology platform can be achieved more efficiently through adoption of a synthetic biology approach. Significance Statement Chlamydomonas reinhardtii offers potential as a host for the production of high value compounds for industrial biotechnology. Synthetic biology provides a mechanism to generate generic, well characterised tools for application in the rational genetic manipulation of organisms: if synthetic biology principles were adopted for manipulation of C. reinhardtii, development of this microalga as an industrial biotechnology platform would be expedited.
Abstract.
Helliwell KE, Collins S, Kazamia E, Purton S, Wheeler GL, Smith AG (2015). Fundamental shift in vitamin B<inf>12</inf> eco-physiology of a model alga demonstrated by experimental evolution.
ISME Journal,
9(6), 1446-1455.
Abstract:
Fundamental shift in vitamin B12 eco-physiology of a model alga demonstrated by experimental evolution
A widespread and complex distribution of vitamin requirements exists over the entire tree of life, with many species having evolved vitamin dependence, both within and between different lineages. Vitamin availability has been proposed to drive selection for vitamin dependence, in a process that links an organism's metabolism to the environment, but this has never been demonstrated directly. Moreover, understanding the physiological processes and evolutionary dynamics that influence metabolic demand for these important micronutrients has significant implications in terms of nutrient acquisition and, in microbial organisms, can affect community composition and metabolic exchange between coexisting species. Here we investigate the origins of vitamin dependence, using an experimental evolution approach with the vitamin B 12 -independent model green alga Chlamydomonas reinhardtii. In fewer than 500 generations of growth in the presence of vitamin B 12, we observe the evolution of a B 12 -dependent clone that rapidly displaces its ancestor. Genetic characterization of this line reveals a type-II Gulliver-related transposable element integrated into the B 12 -independent methionine synthase gene (METE), knocking out gene function and fundamentally altering the physiology of the alga.
Abstract.
2014
Helliwell KE, Scaife MA, Sasso S, Araujo APU, Purton S, Smith AG (2014). Unraveling vitamin B12-responsive gene regulation in algae.
Plant Physiology,
165(1), 388-397.
Abstract:
Unraveling vitamin B12-responsive gene regulation in algae
Photosynthetic microalgae play a vital role in primary productivity and biogeochemical cycling in both marine and freshwater systems across the globe. However, the growth of these cosmopolitan organisms depends on the bioavailability of nutrients such as vitamins. Approximately one-half of all microalgal species requires vitamin B12 as a growth supplement. The major determinant of algal B12 requirements is defined by the isoform of methionine synthase possessed by an alga, such that the presence of the B12-independent methionine synthase (METE) enables growth without this vitamin. Moreover, the widespread but phylogenetically unrelated distribution of B12 auxotrophy across the algal lineages suggests that the METE gene has been lost multiple times in evolution. Given that METE expression is repressed by the presence of B12, prolonged repression by a reliable source of the vitamin could lead to the accumulation of mutations and eventually gene loss. Here, we probe METE gene regulation by B12 and methionine/folate cycle metabolites in both marine and freshwater microalgal species. In addition, we identify a B12-responsive element of Chlamydomonas reinhardtii METE using a reporter gene approach. We show that complete repression of the reporter occurs via a region spanning -574 to -90 bp upstream of the METE start codon. A proteomics study reveals that two other genes (S-Adenosylhomocysteine hydrolase and Serine hydroxymethyltransferase2) involved in the methionine-folate cycle are also repressed by B12 in C. reinhardtii. The strong repressible nature and high sensitivity of the B12-responsive element has promising biotechnological applications as a cost-effective regulatory gene expression tool. © 2014 American Society of Plant Biologists. All rights reserved.
Abstract.
2013
Helliwell KE, Wheeler GL, Smith AG (2013). Widespread decay of vitamin-related pathways: Coincidence or consequence?.
Trends in Genetics,
29(8), 469-478.
Abstract:
Widespread decay of vitamin-related pathways: Coincidence or consequence?
The advent of modern genomics has provided an unparalleled opportunity to consider the gene complement of an organism, and scrutinize metabolic pathways that are no longer active. This approach has led to an increasing number of reports of vitamin-associated pathway deterioration, with many indicating that independent gene-loss events of one or a few key genes have led to vitamin auxotrophy. Nonfunctional unitary pseudogenes belonging to these pathways are found in several species, demonstrating that these are recent evolutionary processes. Here, we examine the commonalities in the cellular roles and metabolism of vitamins that might have led to these losses. The complex pattern of vitamin auxotrophy across the eukaryotic tree of life is intimately connected with the interdependence between organisms. The importance of this process in terms of shaping communities on the one hand, and facilitating symbioses between organisms on the other, is only just beginning to be recognized. © 2013 Elsevier Ltd.
Abstract.
2011
Helliwell KE, Wheeler GL, Leptos KC, Goldstein RE, Smith AG (2011). Insights into the evolution of vitamin B <inf>12</inf> auxotrophy from sequenced algal genomes.
Molecular Biology and Evolution,
28(10), 2921-2933.
Abstract:
Insights into the evolution of vitamin B 12 auxotrophy from sequenced algal genomes
Vitamin B 12 (cobalamin) is a dietary requirement for humans because it is an essential cofactor for two enzymes, methylmalonyl-CoA mutase and methionine synthase (METH). Land plants and fungi neither synthesize or require cobalamin because they do not contain methylmalonyl-CoA mutase, and have an alternative B 12-independent methionine synthase (METE). Within the algal kingdom, approximately half of all microalgal species need the vitamin as a growth supplement, but there is no phylogenetic relationship between these species, suggesting that the auxotrophy arose multiple times through evolution. We set out to determine the underlying cellular mechanisms for this observation by investigating elements of B 12 metabolism in the sequenced genomes of 15 different algal species, with representatives of the red, green, and brown algae, diatoms, and coccolithophores, including both macro-and microalgae, and from marine and freshwater environments. From this analysis, together with growth assays, we found a strong correlation between the absence of a functional METE gene and B 12 auxotrophy. The presence of a METE unitary pseudogene in the B 12-dependent green algae Volvox carteri and Gonium pectorale, relatives of the B 12-independent Chlamydomonas reinhardtii, suggest that B 12 dependence evolved recently in these lineages. In both C. reinhardtii and the diatom Phaeodactylum tricornutum, growth in the presence of cobalamin leads to repression of METE transcription, providing a mechanism for gene loss. Thus varying environmental conditions are likely to have been the reason for the multiple independent origins of B 12 auxotrophy in these organisms. Because the ultimate source of cobalamin is from prokaryotes, the selective loss of METE in different algal lineages will have had important physiological and ecological consequences for these organisms in terms of their dependence on bacteria. © the Author 2011. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All reserved.
Abstract.
2010
Kazamia E, Helliwell KE, Smith AG (2010). Keeping a clear head with vitamin B<inf>12</inf>.
Biochemist,
32(6), 20-24.
Abstract:
Keeping a clear head with vitamin B12
Vitamins are vital organic micronutrients that are required in our diet because they provide essential enzyme cofactors, and animals have dispensed with the ability to synthesize them. Vitamin B12, or cobalamin, is the most complex of the vitamins, and the elucidation of its physiological role, its structure and its biosynthetic pathways have been the subject of impressive scientific endeavours over the years. Vitamin B12 deficiency leads to increased risk of cardiovascular disease, neurological symptoms and, in the most serious cases, pernicious anaemia. Cobalamin is synthesized only by prokaryotes, so we obtain it second-hand by eating other organisms that have accumulated the vitamin in their tissues. The richest dietary sources are liver, dairy products and also algae, many of which are like animals in that they require an exogenous supply of the vitamin for growth. © 2010 the Biochemical Society.
Abstract.