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Comparative transcriptomics of wild-type (UCD 153) and patN-deletion strains (UCD 524) of Nostoc punctiforme ATCC 29133

ABSTRACT: Heterocysts, cells specialized for nitrogen fixation in certain filamentous cyanobacteria, appear singly in a nonrandom spacing pattern along the chain of vegetative cells. A two-stage, biased initiation and competitive resolution model has been proposed to explain the establishment of this spacing pattern. There is substantial evidence that competitive resolution of a subset of cells initiating differentiation occurs by interactions between a self enhancing activator, HetR, and a diffusible inhibitor PatS-5 (RGSGR). Results presented here show that the absence of a unique membrane protein, PatN, in Nostoc punctiforme strain ATCC 29133 leads to a threefold increase in heterocyst frequency and a fourfold decrease in the vegetative cell interval between heterocysts. A PatN-GFP translational fusion shows a pattern of biased inheritance in daughter vegetative cells of ammonium-grown cultures. Inactivation of another heterocyst patterning gene, patA, is epistatic to inactivation of patN, and transcription of patA increases in a patN- deletion strain, implying that patN may function by modulating levels of patA. The presence of PatN is hypothesized to decrease the competency of a vegetative cell to initiate heterocyst differentiation, and the cellular concentration of PatN is dependent on cell division that results in cells transiently depleted of PatN. We suggest that biased inheritance of cell-fate determinants is a phylogenetic domain- spanning paradigm in the development of biological patterns.

ORGANISM(S): Nostoc punctiforme PCC 73102

PROVIDER: GSE40250 | GEO | 2012/10/01

SECONDARY ACCESSION(S): PRJNA173341

REPOSITORIES: GEO

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Comparative transcriptomics of wild-type (UCD 153) and patN-deletion strains (UCD 524) of Nostoc punctiforme ATCC 29133

Project description:Heterocysts, cells specialized for nitrogen fixation in certain filamentous cyanobacteria, appear singly in a nonrandom spacing pattern along the chain of vegetative cells. A two-stage, biased initiation and competitive resolution model has been proposed to explain the establishment of this spacing pattern. There is substantial evidence that competitive resolution of a subset of cells initiating differentiation occurs by interactions between a self enhancing activator, HetR, and a diffusible inhibitor PatS-5 (RGSGR). Results presented here show that the absence of a unique membrane protein, PatN, in Nostoc punctiforme strain ATCC 29133 leads to a threefold increase in heterocyst frequency and a fourfold decrease in the vegetative cell interval between heterocysts. A PatN-GFP translational fusion shows a pattern of biased inheritance in daughter vegetative cells of ammonium-grown cultures. Inactivation of another heterocyst patterning gene, patA, is epistatic to inactivation of patN, and transcription of patA increases in a patN- deletion strain, implying that patN may function by modulating levels of patA. The presence of PatN is hypothesized to decrease the competency of a vegetative cell to initiate heterocyst differentiation, and the cellular concentration of PatN is dependent on cell division that results in cells transiently depleted of PatN. We suggest that biased inheritance of cell-fate determinants is a phylogenetic domain- spanning paradigm in the development of biological patterns. Change in gene expression for a wild-type (UCD 153) and patN-deletion strain, (UCD 524) of Nostoc punctiforme ATCC 29133 over the time course of heterocyst developments. Total RNA from 3 biological replicates at each time point from 0 to 120 hours after removal of combined nitrogen (Nitrogen step-down) was converted to cDNA, dye-labled and hybridized to nimblegen 12x135k array slides.

2012-10-01 | E-GEOD-40250 | biostudies-arrayexpress

Nitrogen-regulated antisense transcription in the adaptation to nitrogen deficiency in Nostoc sp. PCC 7120

Project description:Transcriptomic analyses using high-throughput methods have revealed abundant antisense transcription in bacteria. Most frequently, antisense transcription is due to the overlap of mRNAs with long 5’ regions or 3’ ends that extend beyond the coding sequence. In addition, antisense RNAs that do not contain any coding sequence are also observed. Nostoc sp. PCC 7120 is a filamentous cyanobacterium that, under nitrogen limitation, behaves as a multicellular organism with division of labor among two different cell types that depend on each other, the vegetative CO2-fixing cells and the nitrogen-fixing heterocysts. Differentiation of heterocysts depends on the global nitrogen regulator NtcA and requires the specific regulator HetR. To identify antisense RNAs potentially involved in heterocyst differentiation we performed an RNA-Seq analysis of cells subjected to nitrogen limitation (either at 9 or 24 hours after nitrogen removal) and analyzed the results in combination with a genome-wide set of nitrogen-regulated transcriptional start sites and a prediction of transcriptional terminators. Our analysis resulted in the definition of a transcriptional map including more than 4,000 transcripts, 65% of them in antisense orientation to other transcripts. In addition to overlapping mRNAs we identified nitrogen-regulated non-coding antisense RNAs transcribed from NtcA-dependent or HetR-dependent promoters.

2023-06-27 | GSE212705 | GEO

Cell-specific gene expression in Anabaena variabilis grown phototrophically, mixotrophically, and heterotrophically

2013-11-01 | GSE46076 | GEO

A new world of RNA-binding proteins in heterocyst-forming cyanobacteria.

Project description:RNA-binding proteins (RBPs) are central components of gene regulatory networks and RNA metabolism. The differentiation of nitrogen-fixing heterocysts in some filamentous cyanobacteria and their cooperation with vegetative cells in the same filament is one of the evolutionarily oldest cases of true multicellularity. Nostoc sp. PCC 7120 (Nostoc) is a model organism for filamentous cyanobacteria that differentiate heterocysts. Although hundreds of noncoding transcripts may be involved in this process, the RBPs that could cooperate with them are poorly characterized. We have used R-DeeP/GradR, a technique based on the differential sedimentation of RNA-protein complexes after RNase treatment in density gradients. All macromolecular complexes in the cell are fractionated by a sucrose gradient followed by high-resolution proteomics. We have compared gradients of RNase-treated with untreated samples in triplicates. If a protein was part of an RNA-protein complex, the complex should be disassembled after the RNase treatment and a shift of the protein position in the gradient should be observed (RNA-dependent proteins). Our approach yielded 333 RNA-dependent proteins. The results were compared to a bioinformatic prediction of RBPs and biochemical validation. We validated in vivo the RNA-binding capacity of 6 new RBPs. Some of these proteins were previously known to be involved in essential biological processes such as photosynthesis or heterocyst differentiation, but their association with RNA was previously unknown. In other cases, the function of the validated candidates was unknown. A knock-out strain of alr1700, a gene encoding a validated RNA-binding protein of unknown function, showed a deregulation of heterocyst differentiation, confirming the role of RBPs in regulating this cell differentiation process.

2024-11-08 | PXD050404 | Pride

Time course of nitrogen-responsive transcriptomes in Nostoc sp. PCC7120

Project description:In order to analyze the responses of a heterocystous cyanobacterium to combined-nitrogen depletion/addition, we have used customized microarrays to identify genes potentially involved in heterocysts differentiation by comparing the responses in the control strain and a hetR mutant unable to differentiate heterocysts. RNA samples were isolated from cells growing in the presence of combined nitrogen (ammonium) or after 6, 8, 12 or 24 h of nitrogen deprivation. Additionally, RNA was isolated from cells growing in the absence of combined nitrogen or 8 h after addition of ammonium

2019-05-07 | GSE120377 | GEO

Unbalancing FurB (Zur)-mediated homeostasis in Anabaena sp. PCC7120: Consequences on heterocyst development and biofilm formation

Project description:Here, we report the comparison of transcriptomes of Anabaena sp. PCC7120 and the FurB(Zur) deletion derivative strain (MN38). Anabaena sp PCC7120 is a cyanobacterium that differentiates specialized nitrogen-fixing cells called heterocysts and that is capable of forming biofilms. Our data showed that the deletion of FurB negativily affected the heterocyst development and the biofilm formation. In addition, the RNA-seq data together with gel retardation assays unveiled that FurB is directly involved in the regulation of several genes related to heterocyst development and biofilm formation and other novel functions different from the ones related to the canonical Zur regulon.

2023-09-14 | E-MTAB-12347 | biostudies-arrayexpress

R-DeeP/TripepSVM identifies the RNA-binding OB-fold-like protein PatR as regulator of heterocyst patterning

Project description:RNA-binding proteins (RBPs) are central components of gene regulatory networks. The differentiation of heterocysts in filamentous cyanobacteria is an example of cell differentiation in prokaryotes. Although multiple non-coding transcripts are involved in this process, no RBPs have been implicated thus far. Here we used quantitative mass spectrometry to analyze the differential fractionation of RNA-protein complexes after RNase treatment in density gradients yielding 333 RNA-associated proteins, while a bioinformatic prediction yielded 311 RBP candidates in Nostoc sp. PCC 7120. We validated in vivo the RNA-binding capacity of 6 RBP candidates. Alr1700 is an RBP with two OB-fold domains that is differentially expressed in heterocysts and interacts with non-coding regulatory RNAs. Deletion of alr1700 led to complete deregulation of the cell differentiation process, a striking increase in the number of heterocyst-like cells, and was ultimately lethal in the absence of combined nitrogen. These observations characterize this RBP as a master regulator of the heterocyst patterning and differentiation process, leading us to rename Alr1700 to PatR.

2024-12-20 | GSE280056 | GEO

Cell-specific gene expression in Anabaena variabilis grown phototrophically, mixotrophically, and heterotrophically

Project description:For the filamentous cyanobacterium Anabaena variabilis to grow without combined nitrogen, certain cells differentiate into heterocysts that fix N2, while vegetative cells perform photosynthesis. Much remains unknown on how heterocysts differ from vegetative cells in terms of carbon and energy metabolisms. Microarrays were used to investigate gene transcription patterns in vegetative cells, heterocysts, and filaments of N2-fixing phototrophic, mixotrophic, and heterotrophic cultures. Hybridizations used NimbleGen expression array chips (Product no. A4385-00-01, platform accession no GPL15883) designed against the 5,657 ORFs encoded in the A. variabilis genome (GenBank accession no. CP000117). Each ORF was represented by seventeen 60-mer oligonucleotides. Each oligonucleotide was present in four internal replicates. The twenty-seven microarray data files were normalized against each other. Expression array data were analyzed using ArrayStar 3.0 (DNASTAR, Madison, WI).

2013-11-01 | E-GEOD-46076 | biostudies-arrayexpress

RNA-seq of Nostoc sp. PCC 7120 (Anabaena sp. PCC 7120) wild-type strain compared to a FurC-overexpressing derivative strain grown in BG11 and after 48 hours of nitrogen-step down.

Project description:Here, we report the comparison of transcriptomes of Anabaena sp. PCC7120 and a FurC-overexpressing derivative strain grown under standard conditions (BG11) and after 48 hours of nitrogen step-down (BG110). Anabaena sp PCC7120 is a cyanobacterium that differentiates specialized nitrogen-fixing cells called heterocysts. Our data suggests that FurC directly controls the regulation of heterocyst differentiation and nitrogen fixation in this cyanobacterium. In addition, we found that FurC is also clearly involved in the regulation of several genes belonging to different functional categories, such as iron metabolism, photosynthesis and regulatory functions.

2020-04-28 | E-MTAB-8996 | biostudies-arrayexpress

Protein interaction network reveals the biofunctional difference between vegetative cell and heterocyst in multicellular cyanobacteria

Project description:Protein interaction network reveals the biofunctional difference between vegetative cell and heterocyst in multicellular cyanobacteria

2022-04-26 | PXD025312 |

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