RNA-seq of Nostoc sp. PCC 7120 (Anabaena sp. PCC 7120) wild-type strain, hetZ mutant and hetP mutant
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ABSTRACT: Total RNA was extracted from Anabaena 7120, hetZ mutant and hetP mutant at 24 h after nitrogen stepdown with two independent biological replicates. To maintain the integrity of coding RNAs, total RNA were sequenced without ribosomal RNA elimination. Strand-specific RNA-Seq libraries were prepared and sequenced using the Illumina HiSeq 2500 sequencing instrument to generate paired-end reads with length of 125 bp.
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.
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.
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).
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).
Project description:The whole regulon of the LTTR All3953 was determined at 3 h after Ci deficiency in the cyanobacterium Anabaena sp. PCC 7120 by ChIP-Seq analysis. A TAP-tagged version of the protein was used for the chromatin immunoprecipitation. A total of 142 peaks were found, mainly located in the chromosome of Anabaena.
Project description:The CRP-family transcription factor NtcA, universally found in cyanobacteria, was initially discovered as a regulator operating N control. It responds to the N regime signaled by the internal 2-oxoglutarate levels, an indicator of the C to N balance of the cells. NtcA-activated canonical promoters bear an NtcA-consensus binding site (GTAN8TAC) centered at about 41.5 nucleotides upstream from the transcription start point. In this study, we have used chromatin immunoprecipitation followed by high-throughput sequencing to identify the whole regulon of NtcA in cells of the filamentous, heterocyst-forming cyanobacterium Anabaena sp. PCC 7120 after the withdrawal of combined N. NtcA has been found to bind to 2,424 target regions in the genome of Anabaena, which have been ascribed to 2,153 genes. Interestingly, only a small proportion of those are involved in N assimilation and metabolism, and 65 % of the target regions were located intragenically. The NtcA regulon identified here constitutes the largest bacterial regulon described to date. Our results show that NtcA has a much wider role in the physiology of the cell than it has been previously thought, acting both as a global transcriptional regulator and possibly also as a factor influencing the superstructure of chromosome (and plasmids). Cells of Anabaena sp. PCC 7120 subjected to N-withdrawal for 3 h were used to perform chromatin immunoprecipitation with anti-NtcA antibody. The immunoprecipitated material was sequenced. Three ChIPs were performed from two independent sets of Anabaena cells. A sample of total DNA (not subjected to immunoprecipitation) was used as a control (Input sample).
Project description:we employed a label-free quantitative proteomic strategy to systematically investigate the cellular response to nitrogen deficiency in Anabaena 7120 at different time points