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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

Transcriptome analysis of a N. punctiforme sigG mutant

2015-09-01 | GSE61442 | GEO

Transcriptome analysis of a N. punctiforme sigG mutant

Project description:Nostoc punctiforme is a filamentous cyanobacterium capable if differentiating normal vegetative cells into nitrogen fixing heterocysts and spore-like akinetes. Here the effects of a sigma factor (sigG) mutation on the transcriptome was analyzed under 1) normal vegetative growth, and 2) following akinete induction. A zwf strain grown with fructose differentiates into akinetes following transfer to darkness. A DNA microarray was used to compare gene expression from the zwf strain to a zwf sigG double mutant two days following akinte induction by darkness. Gene expression in a zwf mutant strain was compared to a zwf sigG double mutant strain under normal vegetative growth and at two days following akinete induction. Each experiment used three array slides with two biological replicates. A dye swap was used for one biological replicate.

2015-09-01 | E-GEOD-61442 | biostudies-arrayexpress

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

2013-11-01 | GSE46076 | GEO

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

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

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

Deep sequencing of HetR-bound DNA reveals novel HetR targets in Anabaena sp. strain PCC 7120

Project description:We employed chromatin pull-down and deep sequencing to globally identify HetR DNA targets in vivo at 6 hours after fixed-nitrogen deprivation. We identified novel DNA binding targets of tagged HetR-6xHis and defined a consensus HetR binding site from these HetR target sequences. Chromatin pull down of hetR mutant strain UHM103 carrying pAM4375, which expresses HetR-6xHis, and a wild-type control. One dataset was collected.

2013-10-31 | E-GEOD-51886 | biostudies-arrayexpress

Deep sequencing of HetR-bound DNA reveals novel HetR targets in Anabaena sp. strain PCC 7120

2013-10-31 | GSE51886 | GEO

Transcriptional Regulation of Anabaena 7120 response to Nitrogen Deprivation

Project description:Here, we report the transcriptome of Anabaena sp. strain 7120, a cyanobacterium that forms specialized nitrogen-fixing cells called heterocysts. Our data suggests that cyanobacteria frequently have more complex transcripts than thought, with large 5' UTRs, numerous antisense transcripts, and multiple transcriptional start sites or processing sites. Four samples of total filament RNA were sequenced with Illumina 40bp reads using directional RNA sequencing (see the Illumina small RNA prep protocol). The samples are 0hr (vegetative cells grown in the presence of ammonia) and 6hr, 12hr, and 21hr cells (after nitrogen step down).

2011-06-22 | E-GEOD-26633 | biostudies-arrayexpress

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