A protein is involved in accessibility of the inhibitor acetazolamide to the carbonic anhydrase(s) in the cyanobacterium Synechocystis PCC 6803

Beuf, Laurent; Bédu, Sylvie; Cami, B.; Joset, F.

doi:10.1007/bf00020230

Cited by 5 publications

(8 citation statements)

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“…A1543 branched more closely to Sll1910 (Fig. 7a), a protein previously reported to confer sensitivity to the carbonic anhydrase inhibitor, acetazolamide (Beuf et al 1995; Bedu et al 1990). In contrast to A0574 , we were able to replace the A1543 gene with a Sp R cassette (Fig.…”

Section: Resultsmentioning

confidence: 79%

“…The ortholog of A0574, Sll1290 in PCC6803 (67% identity) was recently shown to have RNase II activity (Matos et al 2012); thus, it is possible that A0574 also shares this activity. A mutant of the A1543 ortholog of in PCC6803, A1910, was isolated in a screen for resistance to the carbonic anhydrase inhibitor acetazolamide and renamed Zam (Beuf et al 1995; Bedu et al 1990). However, the mechanism for the observed resistance to acetazolamide was not elucidated.…”

Section: Discussionmentioning

confidence: 99%

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Genetic and genomic analysis of RNases in model cyanobacteria

2015

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Cyanobacteria are diverse photosynthetic microbes with the ability to convert CO2 into useful products. However, metabolic engineering of cyanobacteria remains challenging because of the limited resources for modifying the expression of endogenous and exogenous biochemical pathways. Fine-tuned control of protein production will be critical to optimize the biological conversion of CO2 into desirable molecules. Messenger RNA (mRNA) are labile intermediates that play critical roles in determining the translation rate and steady state protein concentrations in the cell. The majority of studies on mRNA turnover have focused on the model heterotrophic bacteria Escherichia coli and Bacillus subtilis. These studies have elucidated many RNA modifying and processing enzymes and have highlighted the differences between these Gram-negative and Gram-positive bacteria, respectively. In contrast, much less is known about mRNA turnover in cyanobacteria. We generated a compendium of the major ribonucleases (RNases) and provide an in-depth analysis of RNase III-like enzymes in commonly studied and diverse cyanobacteria. Furthermore, using targeted gene deletion, we genetically dissected the RNases in Synechococcus sp. PCC 7002, one of the fastest growing and industrially attractive cyanobacterial strains. We found that all three cyanobacterial homologs of RNase III and a member of the RNase II/R family are not essential under standard laboratory conditions, while homologs of RNase E/G, RNase J1/J2, PNPase, and a different member of the RNase II/R family appear to be essential for growth. This work will enhance our understanding of native control of gene expression and will facilitate the development of an RNA-based toolkit for metabolic engineering in cyanobacteria.

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Section: Resultsmentioning

confidence: 79%

Section: Discussionmentioning

confidence: 99%

Genetic and genomic analysis of RNases in model cyanobacteria

2015

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“…In recent years, it has been reported that SSD1 has a weak but significant similarity with dis3 ϩ of Schizosaccharomyces pombe (6,13), DSS1 of S. cerevisiae (14), vacB of Shigella flexneri (15), cyt4 of Neurospora crassa (16), zam of Synechocytosis PCC 6803 (17), and rnb of Escherichia coli (18). Some of these genes are known, or implied, to be involved in the modification of RNAs: 1) cyt4 is required for the mitochondrial rRNA splicing and processing reaction; 2) DSS1 is a multicopy suppressor of the disruptant of SUV3 encoding a putative RNA helicase-like protein; 3) the vacB mutation reduces the level of the virulence antigens, IpaB, IpaC, IpaD, and VirG, at the post-transcriptional level; and 4) the RNase II encoded by rnb has a 3Ј-to-5Ј exoribonuclease activity.…”

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confidence: 99%

Ssd1p of Saccharomyces cerevisiae Associates with RNA

Uesono

Toh‐e

Kikuchi

1997

Journal of Biological Chemistry

102

123

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The SSD1 gene has been isolated as a single copy suppressor of many mutants, such as sit4, slk1/bck1, pde2, and rpc31, in the yeast Saccharomyces cerevisiae. Ssd1p has domains showing weak but significant homology with RNase II-related proteins, Cyt4p, Dss1p, VacB, and RNase II, which are involved in the modification of RNA. We found that Ssd1p had the ability to bind RNA, preferably poly(rA), as well as single-stranded DNA. Interestingly, the most conserved domain among the RNase II-related proteins was not necessary for interaction with RNA. Indirect immunofluorescence staining with anti-Ssd1p antibody revealed that Ssd1p was detected mainly in the cytoplasm. Furthermore, sucrose gradient sedimentation analysis demonstrated that Ssd1p was not cofractionated with polyribosomes, suggesting that Ssd1p is not particularly bound to a translationally active subpopulation of mRNA in the cytoplasm.Cellular RNAs do not exist as a free form but as an RNAprotein complex. The proteins that directly associate with RNA are thought to play important roles in the regulation of gene expression at the post-transcriptional level (1, 2). In eukaryotic cells, proteins that bind to RNA polymerase II transcripts include both heterogeneous nuclear RNA-binding proteins and cytoplasmic mRNA-binding proteins. Heterogeneous nuclear RNA-binding proteins bind pre-mRNAs and are associated with them during the processing events required for the formation of mature mRNA (1). Once mRNAs are transported to the cytoplasm, they form cytoplasmic mRNA-binding protein complexes (2). Cytoplasmic mRNA-binding proteins seem to regulate translation, localization, or stability of mRNA (3). At present, many RNA-binding proteins have been isolated and characterized (4, 5), but their functions have not been fully understood.In Saccharomyces cerevisiae, the SSD1 gene has been first characterized to suppress the sit4 mutation defective in a protein phosphatase subunit (6). Not only in this case, but also in many other cases, SSD1 has been isolated as a single copy suppressor of mutation defective in RPC31 encoding a subunit of RNA polymerase III (7), in PDE2 encoding the cyclic AMP phosphodiesterase (8), in BCK1 encoding mitogen-activated protein kinase kinase kinase (9), in MPK1 encoding mitogenactivated protein kinase (10), or in G 1 cyclin (11). These reports indicate that SSD1 is involved in many systems. Sutton et al.also reported that there are two alleles of the SSD1 gene; one is called ssd1-d (dead) and the other is called SSD1-V (viable). They described that SSD1-V could suppress the double mutations of ssd1-d and sit4 (6). We have also isolated the SSD1 gene as the MCS1 gene involved in stable maintenance of the minichromosome (12). The SSD1/MCS1 gene product was detected as a ϳ160-kDa protein in certain wild type strains bearing SSD1-V, such as KA31 or RAY-3A, whereas a protein of this size was not detected in another wild type strain bearing ssd1-d, such as YPH499 (7). These findings indicate that SSD1-V is simply a wild type gene and ssd1-d is a ...

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“…PCC 6803 (hereafter PCC 6803) [ 3 ]. Either a deletion or an insertion mutation in zam confers resistance to lethal concentrations of the carbonic anhydrase inhibitor acetazolamide to PCC 6803 [ 4 ]. Zam was also identified as a redox-responsive protein in both PCC 6803 and Synechococcus sp.…”

Section: Introductionmentioning

confidence: 99%

“…PCC 7002 (hereafter PCC 7002) in two separate redox-proteomics experiments [ 5 , 6 ]. It was initially presumed that zam encoded a transporter which affected the accessibility of acetazolamide to one or more of the cellular carbonic anhydrases; however, multiple sequence alignments show that Zam is actually a member of the RNase II/R family [ 4 , 7 ]. Redox-based control of RNA modulating enzymes is not unprecedented in cyanobacteria.…”

Section: Introductionmentioning

confidence: 99%

Zam Is a Redox-Regulated Member of the RNB-Family Required for Optimal Photosynthesis in Cyanobacteria

et al. 2022

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The zam gene mediating resistance to acetazolamide in cyanobacteria was discovered thirty years ago during a drug tolerance screen. We use phylogenetics to show that Zam proteins are distributed across cyanobacteria and that they form their own unique clade of the ribonuclease II/R (RNB) family. Despite being RNB family members, multiple sequence alignments reveal that Zam proteins lack conservation and exhibit extreme degeneracy in the canonical active site—raising questions about their cellular function(s). Several known phenotypes arise from the deletion of zam, including drug resistance, slower growth, and altered pigmentation. Using room-temperature and low-temperature fluorescence and absorption spectroscopy, we show that deletion of zam results in decreased phycocyanin synthesis rates, altered PSI:PSII ratios, and an increase in coupling between the phycobilisome and PSII. Conserved cysteines within Zam are identified and assayed for function using in vitro and in vivo methods. We show that these cysteines are essential for Zam function, with mutation of either residue to serine causing phenotypes identical to the deletion of Zam. Redox regulation of Zam activity based on the reversible oxidation-reduction of a disulfide bond involving these cysteine residues could provide a mechanism to integrate the ‘central dogma’ with photosynthesis in cyanobacteria.

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A protein is involved in accessibility of the inhibitor acetazolamide to the carbonic anhydrase(s) in the cyanobacterium Synechocystis PCC 6803

Cited by 5 publications

References 32 publications

Genetic and genomic analysis of RNases in model cyanobacteria

Genetic and genomic analysis of RNases in model cyanobacteria

Ssd1p of Saccharomyces cerevisiae Associates with RNA

Zam Is a Redox-Regulated Member of the RNB-Family Required for Optimal Photosynthesis in Cyanobacteria

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