Illumination stimulates cAMP receptor protein-dependent transcriptional activation from regulatory regions containing class I and class II promoter elements in Synechocystis sp. PCC 6803

Hedger, Jennifer; Holmquist, Peter C.; Leigh, Kimberly A.; Saraff, Kumuda; Pomykal, Christina; Summers, Michael L.

doi:10.1099/mic.0.028035-0

Cited by 10 publications

(15 citation statements)

References 46 publications

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“…A TSS 206 nt upstream of murF was mapped, which indicates that this is indeed the case. This TSS has very recently been confirmed by another group (Hedger et al , 2009). Interestingly, we found the expression level of SyR7 more than 20 times higher than that of the murF mRNA under three different conditions.…”

Section: Resultssupporting

confidence: 72%

“…However, on a shift to HL, the SyR7/ murF ratio declined dramatically to ∼1 (Figure 6A). This change may be in part because of activation of the P1 promoter activated by the cAMP‐dependent transcription factor SyCRP1 (Hedger et al , 2009), contributing to a slight increase in murF mRNA concentrations, by a factor of 2.34±0.56 (Figure 6A). The light effect on the SyR7 steady state level was, however, much more pronounced as it declined to only 15% of its initial value (Figure 6A).…”

Section: Resultsmentioning

confidence: 99%

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Evidence for a major role of antisense RNAs in cyanobacterial gene regulation

Georg

Voß

Scholz

et al. 2009

Molecular Systems Biology

157

188

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Information on the numbers and functions of naturally occurring antisense RNAs (asRNAs) in eubacteria has thus far remained incomplete. Here, we screened the model cyanobacterium Synechocystis sp. PCC 6803 for asRNAs using four different methods. In the final data set, the number of known noncoding RNAs rose from 6 earlier identified to 60 and of asRNAs from 1 to 73 (28 were verified using at least three methods). Among these, there are many asRNAs to housekeeping, regulatory or metabolic genes, as well as to genes encoding electron transport proteins. Transferring cultures to high light, carbon-limited conditions or darkness influenced the expression levels of several asRNAs, suggesting their functional relevance. Examples include the asRNA to rpl1 , which accumulates in a light-dependent manner and may be required for processing the L11 r-operon and the SyR7 noncoding RNA, which is antisense to the murF 5′ UTR, possibly modulating murein biosynthesis. Extrapolated to the whole genome, ∼10% of all genes in Synechocystis are influenced by asRNAs. Thus, chromosomally encoded asRNAs may have an important function in eubacterial regulatory networks.

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Section: Resultssupporting

confidence: 72%

Section: Resultsmentioning

confidence: 99%

Evidence for a major role of antisense RNAs in cyanobacterial gene regulation

Georg

Voß

Scholz

et al. 2009

Molecular Systems Biology

157

188

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“…For initiation of transcription the Crp has to be on the same side of the DNA molecule as the RNAP, which limits the spacing from the middle of the –10 sigma factor-binding site to the middle of the Crp site in intervals of 10.5-bp α-helical turns (45). By definition class I Crp dependent promoters have five turns or more.…”

Section: Discussionmentioning

confidence: 99%

Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology

Huang¹,

Camsund²,

Lindblad³

et al. 2010

Nucleic Acids Research

312

401

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Cyanobacteria are suitable for sustainable, solar-powered biotechnological applications. Synthetic biology connects biology with computational design and an engineering perspective, but requires efficient tools and information about the function of biological parts and systems. To enable the development of cyanobacterial Synthetic Biology, several molecular tools were developed and characterized: (i) a broad-host-range BioBrick shuttle vector, pPMQAK1, was constructed and confirmed to replicate in Escherichia coli and three different cyanobacterial strains. (ii) The fluorescent proteins Cerulean, GFPmut3B and EYFP have been demonstrated to work as reporter proteins in cyanobacteria, in spite of the strong background of photosynthetic pigments. (iii) Several promoters, like PrnpB and variants of PrbcL, and a version of the promoter Ptrc with two operators for enhanced repression, were developed and characterized in Synechocystis sp. strain PCC6803. (iv) It was shown that a system for targeted protein degradation, which is needed to enable dynamic expression studies, is working in Synechocystis sp. strain PCC6803. The pPMQAK1 shuttle vector allows the use of the growing numbers of BioBrick parts in many prokaryotes, and the other tools herein implemented facilitate the development of new parts and systems in cyanobacteria.

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“…In class III promoters (for example, the acsP2 promoter), CRP interacts with DNA at multiple sites in combination with additional proteins (45). In Synechocystis 6803, Sycrp1 binds in vitro to proposed CRP target sites upstream of slr1351 (murF), sll1874 (chlAII), sll1708 (narL), slr0442, and sll1268 (46), and the genes slr1351, sll1874, and slr0442 were shown to be targets regulated by Sycrp1 (47). The transcriptional profiles of cccS and cccP appeared to be contradictory between Sycrp1-and Sycrp2-disrupted mutants.…”

Section: Discussionmentioning

confidence: 99%

Sycrp2 Is Essential for Twitching Motility in the Cyanobacterium Synechocystis sp. Strain PCC 6803

Song

Zang

et al. 2018

J Bacteriol

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Two cAMP receptor proteins (CRPs), Sycrp1 (encoded by ) and Sycrp2 (encoded by), exist in the cyanobacterium sp. strain PCC 6803. Previous studies have demonstrated that Sycrp1 has binding affinity for cAMP and is involved in motility by regulating the formation of pili. However, the function of Sycrp2 remains unknown. Here, we report that disruption results in the loss of motility of sp. PCC 6803, and that the phenotype can be recovered by reintroducing the gene into the genome of -disrupted mutants. Electron microscopy showed that the-disrupted mutant lost the pilus apparatus on the cell surface, resulting in a lack of cell motility. Furthermore, the transcript level of the operon (essential for cell motility and regulated by the cAMP receptor protein Sycrp1) was markedly decreased in-disrupted mutants compared with the wild-type strain. Moreover, yeast two-hybrid analysis and a pulldown assay demonstrated that Sycrp2 interacted with Sycrp1 to form a heterodimer and that Sycrp1 and Sycrp2 interacted with themselves to form homodimers. Gel mobility shift assays revealed that Sycrp1 specifically binds to the upstream region of Together, these findings indicate that in sp. PCC 6803, Sycrp2 regulates the formation of pili and cell motility by interacting with Sycrp1. cAMP receptor proteins (CRPs) are widely distributed in cyanobacteria and play important roles in regulating gene expression. Although many cyanobacterial species have two cAMP receptor-like proteins, the functional links between them are unknown. Here, we found that Sycrp2 in the cyanobacterium sp. strain PCC 6803 is essential for twitching motility and that it interacts with Sycrp1, a known cAMP receptor protein involved with twitching motility. Our findings indicate that the two cAMP receptor-like proteins in cyanobacteria do not have functional redundancy but rather work together.

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Illumination stimulates cAMP receptor protein-dependent transcriptional activation from regulatory regions containing class I and class II promoter elements in Synechocystis sp. PCC 6803

Cited by 10 publications

References 46 publications

Evidence for a major role of antisense RNAs in cyanobacterial gene regulation

Evidence for a major role of antisense RNAs in cyanobacterial gene regulation

Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology

Sycrp2 Is Essential for Twitching Motility in the Cyanobacterium Synechocystis sp. Strain PCC 6803

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