41 research outputs found
The role of the cytoskeletal proteins MreB and FtsZ in multicellular cyanobacteria
Multiseriate and true-branching cyanobacteria are at the peak of prokaryotic morphological complexity. However, little is known about the mechanisms governing multiplanar cell division and morphogenesis. Here, we study the function of the prokaryotic cytoskeletal proteins, MreB and FtsZ in Fischerella muscicola PCC 7414 and Chlorogloeopsis fritschii PCC 6912. Vancomycin and HADA labeling revealed a mixed apical, septal, and lateral trichome growth mode in F. muscicola, whereas C. fritschii exhibits septal growth. In all morphotypes from both species, MreB forms either linear filaments or filamentous strings and can interact with FtsZ. Furthermore, multiplanar cell division in F. muscicola likely depends on FtsZ dosage. Our results lay the groundwork for future studies on cytoskeletal proteins in morphologically complex cyanobacteria
Structural Determinants and Their Role in Cyanobacterial Morphogenesis
Cells have to erect and sustain an organized and dynamically adaptable structure for an efficient mode of operation that allows drastic morphological changes during cell growth and cell division. These manifold tasks are complied by the so-called cytoskeleton and its associated proteins. In bacteria, FtsZ and MreB, the bacterial homologs to tubulin and actin, respectively, as well as coiled-coil-rich proteins of intermediate filament (IF)-like function to fulfil these tasks. Despite generally being characterized as Gram-negative, cyanobacteria have a remarkably thick peptidoglycan layer and possess Gram-positive-specific cell division proteins such as SepF and DivIVA-like proteins, besides Gram-negative and cyanobacterial-specific cell division proteins like MinE, SepI, ZipN (Ftn2) and ZipS (Ftn6). The diversity of cellular morphologies and cell growth strategies in cyanobacteria could therefore be the result of additional unidentified structural determinants such as cytoskeletal proteins. In this article, we review the current advances in the understanding of the cyanobacterial cell shape, cell division and cell growth
Two novel heteropolymerâforming proteins maintain the multicellular shape of the cyanobacterium Anabaena sp. PCC 7120
Polymerizing and filament-forming proteins are instrumental for numerous cellular processes such as cell division and growth. Their function in stabilization and localization of protein complexes and replicons is achieved by a filamentous structure. Known filamentous proteins assemble into homopolymers consisting of single subunits â for example, MreB and FtsZ in bacteria â or heteropolymers that are composed of two subunits, for example, keratin and α/ÎČ tubulin in eukaryotes. Here, we describe two novel coiled-coil-rich proteins (CCRPs) in the filament-forming cyanobacterium Anabaena sp. PCC 7120 (hereafter Anabaena) that assemble into a heteropolymer and function in the maintenance of the Anabaena multicellular shape (termed trichome). The two CCRPs â Alr4504 and Alr4505 (named ZicK and ZacK) â are strictly interdependent for the assembly of protein filaments in vivo and polymerize nucleotide independently in vitro, similar to known intermediate filament (IF) proteins. A ÎzicKÎzacK double mutant is characterized by a zigzagged cell arrangement and hence a loss of the typical linear Anabaena trichome shape. ZicK and ZacK interact with themselves, with each other, with the elongasome protein MreB, the septal junction protein SepJ and the divisome associate septal protein SepI. Our results suggest that ZicK and ZacK function in cooperation with SepJ and MreB to stabilize the Anabaena trichome and are likely essential for the manifestation of the multicellular shape in Anabaena. Our study reveals the presence of filament-forming IF-like proteins whose function is achieved through the formation of heteropolymers in cyanobacteria
The Smallest Known Genomes of Multicellular and Toxic Cyanobacteria: Comparison, Minimal Gene Sets for Linked Traits and the Evolutionary Implications
Cyanobacterial morphology is diverse, ranging from unicellular spheres or rods to multicellular structures such as colonies and filaments. Multicellular species represent an evolutionary strategy to differentiate and compartmentalize certain metabolic functions for reproduction and nitrogen (N2) fixation into specialized cell types (e.g. akinetes, heterocysts and diazocytes). Only a few filamentous, differentiated cyanobacterial species, with genome sizes over 5 Mb, have been sequenced. We sequenced the genomes of two strains of closely related filamentous cyanobacterial species to yield further insights into the molecular basis of the traits of N2 fixation, filament formation and cell differentiation. Cylindrospermopsis raciborskii CS-505 is a cylindrospermopsin-producing strain from Australia, whereas Raphidiopsis brookii D9 from Brazil synthesizes neurotoxins associated with paralytic shellfish poisoning (PSP). Despite their different morphology, toxin composition and disjunct geographical distribution, these strains form a monophyletic group. With genome sizes of approximately 3.9 (CS-505) and 3.2 (D9) Mb, these are the smallest genomes described for free-living filamentous cyanobacteria. We observed remarkable gene order conservation (synteny) between these genomes despite the difference in repetitive element content, which accounts for most of the genome size difference between them. We show here that the strains share a specific set of 2539 genes with >90% average nucleotide identity. The fact that the CS-505 and D9 genomes are small and streamlined compared to those of other filamentous cyanobacterial species and the lack of the ability for heterocyst formation in strain D9 allowed us to define a core set of genes responsible for each trait in filamentous species. We presume that in strain D9 the ability to form proper heterocysts was secondarily lost together with N2 fixation capacity. Further comparisons to all available cyanobacterial genomes covering almost the entire evolutionary branch revealed a common minimal gene set for each of these cyanobacterial traits
Physiogenomiks von Cylindrospermopsis raciborskii und Raphidiopsis brookii (Cyanobakterien) mit Schwerpunkt auf Evolution, Stickstoff-Kontrolle- und Toxin-Biosynthese
The freshwater toxic cyanobacteria Cylindrospermopsis raciborskii CS-505 and Raphidiopsis brookii D9 belong to the order Nostocales of filamentous, heterocystous (nitrogen fixation) morphotypes. However, Raphidiopsis spp. do not develop heterocyst nor fix N2, contradicting the morphological classification. This thesis presents an integrated study of evolutionary relationships linked to nitrogen (N) metabolism and toxin production in filamentous cyanobacteria. The genome sequencing and comparison of CS-505 and D9 revealed the smallest genomes of filamentous cyanobacteria and an extent of similarity corresponding to conspecific isolates, therefore recommending a taxonomical reclassification of the genera Raphidiopsis. The presence of heterocyst-specific genes and their expression under alternative N-sources in D9, together with differences in genomic plasticity, allowed us to propose divergent evolutionary pathways for both species. Late transcriptional responses were observed in C. raciborskii CS-505 under N-deprivation, contrasting with those of other heterocystous cyanobacteria. The pattern of biosynthetic gene regulation suggested that additional mechanisms of N-control and post-transcriptional mechanisms are involved in toxin production. The role of repetitive elements and transposases in gene regulation may be critical but this aspect requires further investigation
Physiogenomics of Cylindrospermopsis raciborskii and Raphidiopsis brookii (Cyanobacteria) with Emphasis on Evolution, Nitrogen Control and Toxin Biosynthesis
The freshwater toxic cyanobacteria Cylindrospermopsis raciborskii CS-505 and Raphidiopsis brookii D9 belong to the order Nostocales of filamentous, heterocystous (nitrogen fixation) morphotypes. However, Raphidiopsis spp. do not develop heterocyst nor fix N2, contradicting the morphological classification. This thesis presents an integrated study of evolutionary relationships linked to nitrogen (N) metabolism and toxin production in filamentous cyanobacteria. The genome sequencing and comparison of CS-505 and D9 revealed the smallest genomes of filamentous cyanobacteria and an extent of similarity corresponding to conspecific isolates, therefore recommending a taxonomical reclassification of the genera Raphidiopsis. The presence of heterocyst-specific genes and their expression under alternative N-sources in D9, together with differences in genomic plasticity, allowed us to propose divergent evolutionary pathways for both species. Late transcriptional responses were observed in C. raciborskii CS-505 under N-deprivation, contrasting with those of other heterocystous cyanobacteria. The pattern of biosynthetic gene regulation suggested that additional mechanisms of N-control and post-transcriptional mechanisms are involved in toxin production. The role of repetitive elements and transposases in gene regulation may be critical but this aspect requires further investigation
Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering
Impact of Nitrogen Sources on Gene Expression and Toxin Production in the Diazotroph Cylindrospermopsis raciborskii CS-505 and Non-Diazotroph Raphidiopsis brookii D9
Different environmental nitrogen sources play selective roles in the development of cyanobacterial blooms and noxious effects are often exacerbated when toxic cyanobacteria are dominant. Cylindrospermopsis raciborskii CS-505 (heterocystous, nitrogen fixing) and Raphidiopsis brookii D9 (non-N2 fixing) produce the nitrogenous toxins cylindrospermopsin (CYN) and paralytic shellfish toxins (PSTs), respectively. These toxin groups are biosynthesized constitutively by two independent putative gene clusters, whose flanking genes are target for nitrogen (N) regulation. It is not yet known how or if toxin biosynthetic genes are regulated, particularly by N-source dependency. Here we show that binding boxes for NtcA, the master regulator of N metabolism, are located within both gene clusters as potential regulators of toxin biosynthesis. Quantification of intra- and extracellular toxin content in cultures at early stages of growth under nitrate, ammonium, urea and N-free media showed that N-sources influence neither CYN nor PST production. However, CYN and PST profiles were altered under N-free medium resulting in a decrease in the predicted precursor toxins (doCYN and STX, respectively). Reduced STX amounts were also observed under growth in ammonium. Quantification of toxin biosynthesis and transport gene transcripts revealed a constitutive transcription under all tested N-sources. Our data support the hypothesis that PSTs and CYN are constitutive metabolites whose biosynthesis is correlated to cyanobacterial growth rather than directly to specific environmental conditions. Overall, the constant biosynthesis of toxins and expression of the putative toxin-biosynthesis genes supports the usage of qPCR probes in water quality monitoring of toxic cyanobacteria
Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering
Cyanobacteria display a large diversity of cellular forms ranging from unicellular to complex multicellular filaments or aggregates. Species in the group present a wide range of metabolic characteristics including the fixation of atmospheric nitrogen, resistance to extreme environments, production of hydrogen, secondary metabolites and exopolysaccharides. These characteristics led to the growing interest in cyanobacteria across the fields of ecology, evolution, cell biology and biotechnology. The number of available cyanobacterial genome sequences has increased considerably in recent years, with more than 140 fully sequenced genomes to date. Genetic engineering of cyanobacteria is widely applied to the model unicellular strains Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. However the establishment of transformation protocols in many other cyanobacterial strains is challenging. One obstacle to the development of these novel model organisms is that many species have doubling times of 48 h or more, much longer than the bacterial models E. coli or B. subtilis. Furthermore, cyanobacterial defense mechanisms against foreign DNA pose a physical and biochemical barrier to DNA insertion in most strains. Here we review the various barriers to DNA uptake in the context of lateral gene transfer among microbes and the various mechanisms for DNA acquisition within the prokaryotic domain. Understanding the cyanobacterial defense mechanisms is expected to assist in the development and establishment of novel transformation protocols that are specifically suitable for this group