PlmA, a New Member of the GntR Family, Has Plasmid Maintenance Functions in
            <i>Anabaena</i>
            sp. Strain PCC 7120

Lee, Martin H.; Scherer, Michael; Rigali, Sébastien; Golden, James W.

doi:10.1128/jb.185.15.4315-4325.2003

Cited by 63 publications

(68 citation statements)

References 54 publications

(35 reference statements)

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“…For all constructs, the ribosome-binding site and start codon were derived from the promoters used. In all constructs, the inserts were downstream of a transcription terminator on shuttle vectors pAM504, pAM505, and pAM2770 (23,32,36). All plasmid constructs were confirmed by sequencing.…”

Section: Methodsmentioning

confidence: 99%

“…Expression of full-length patS and the patS5 minigene from the rbcL promoter in constructs P rbcL -patS17 (pAM1690) (23) and P rbcL -patS5 (pAM2537) completely inhibited heterocyst formation in both the wild-type strain and the patS null mutant AMC451 (Table 4). P rbcL is strongly expressed in vegetative cells (7,9).…”

Section: Expression Of the Pats5 Minigene In Proheterocysts Failed Tomentioning

confidence: 99%

“…The resulting XhoI-SapI fragment was inserted into pAM2600, resulting in pAM2821, such that patS is translationally fused to six histidine codons. Second, an XhoI-ClaI fragment carrying patS-6His from pAM2821 was inserted into shuttle vector pAM2770 (23), resulting in plasmid pAM2826.…”

Section: Methodsmentioning

confidence: 99%

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patS Minigenes Inhibit Heterocyst Development of Anabaena sp. Strain PCC 7120

Duan

Lee

et al. 2004

J Bacteriol

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The patS gene encodes a small peptide that is required for normal heterocyst pattern formation in the cyanobacterium Anabaena sp. strain PCC 7120. PatS is proposed to control the heterocyst pattern by lateral inhibition. patS minigenes were constructed and expressed by different developmentally regulated promoters to gain further insight into PatS signaling. patS minigenes patS4 to patS8 encode PatS C-terminal 4 (GSGR) to 8 (CDERGSGR) oligopeptides. When expressed by P petE , P patS , or P rbcL promoters, patS5 to patS8 inhibited heterocyst formation but patS4 did not. In contrast to the full-length patS gene, P hepA -patS5 failed to restore a wild-type pattern in a patS null mutant, indicating that PatS-5 cannot function in cell-to-cell signaling if it is expressed in proheterocysts. To establish the location of the PatS receptor, PatS-5 was confined within the cytoplasm as a gfp-patS5 fusion. The green fluorescent protein GFP-PatS-5 fusion protein inhibited heterocyst formation. Similarly, full-length PatS with a C-terminal hexahistidine tag inhibited heterocyst formation. These data indicate that the PatS receptor is located in the cytoplasm, which is consistent with recently published data indicating that HetR is a PatS target. We speculated that overexpression of other Anabaena strain PCC 7120 RGSGR-encoding genes might show heterocyst inhibition activity. In addition to patS and hetN, open reading frame (ORF) all3290 and an unannotated ORF, orf77, encode an RGSGR motif. Overexpression of all3290 and orf77 under the control of the petE promoter inhibited heterocyst formation, indicating that the RGSGR motif can inhibit heterocyst development in a variety of contexts.

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Section: Methodsmentioning

confidence: 99%

Section: Expression Of the Pats5 Minigene In Proheterocysts Failed Tomentioning

confidence: 99%

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patS Minigenes Inhibit Heterocyst Development of Anabaena sp. Strain PCC 7120

Duan

Lee

et al. 2004

J Bacteriol

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“…To date, in addition to AraR, 21 proteins presenting this mosaic modular association in different bacteria have been sequenced. Together they constitute one of the six GntR subfamilies typified by AraR (17,38). However, no functional studies have been reported for these AraR-like proteins (http://www.sanger.ac.uk /Software/Pfam).…”

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confidence: 99%

Functional Domains of theBacillus subtilisTranscription Factor AraR and Identification of Amino Acids Important for Nucleoprotein Complex Assembly and Effector Binding

et al. 2006

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The Bacillus subtilis AraR transcription factor represses at least 13 genes required for the extracellular degradation of arabinose-containing polysaccharides, transport of arabinose, arabinose oligomers, xylose, and galactose, intracellular degradation of arabinose oligomers, and further catabolism of this sugar. AraR exhibits a chimeric organization comprising a small N-terminal DNA-binding domain that contains a winged helix-turn-helix motif similar to that seen with the GntR family and a larger C-terminal domain homologous to that of the LacI/GalR family. Here, a model for AraR was derived based on the known crystal structures of the FadR and PurR regulators from Escherichia coli. We have used random mutagenesis, deletion, and construction of chimeric LexA-AraR fusion proteins to map the functional domains of AraR required for DNA binding, dimerization, and effector binding. Moreover, predictions for the functional role of specific residues were tested by site-directed mutagenesis. In vivo analysis identified particular amino acids required for dimer assembly, formation of the nucleoprotein complex, and composition of the sugar-binding cleft. This work presents a structural framework for the function of AraR and provides insight into the mechanistic mode of action of this modular repressor.The transcription factor AraR controls the expression of several Bacillus subtilis genes encoding enzymes and permeases involved in the degradation of arabinose-containing polysaccharides, uptake of L-arabinose (and possibly arabinose oligomers), xylose, and galactose, and further intracellular catabolism of arabinose and arabinose oligomers. The arabinose (ara) regulon comprises at least 13 genes (Fig. 1), the araAB DLMNPQ-abfA operon (43), the divergently arranged araE/ araR genes (42, 45) located in distinct regions of the B. subtilis chromosome, and the genes abnA and xsa positioned immediately upstream and 23 kb downstream from the operon, respectively (57). The first three genes of the arabinose metabolic operon, araA, araB, and araD, encode the enzymes required for the intracellular conversion of L-arabinose into D-xylulose 5-phosphate, which is further catabolized through the pentose phosphate pathway (41). The function of araL and araM is unknown, and genes araNPQ encode components of an ABC-type transporter most likely involved in the uptake of arabinose oligomers (43, 45). The product of the araE gene is a permease, the main transporter of arabinose into the cell (45), that is also responsible for the uptake of xylose and galactose (14). The araR gene encodes the regulatory protein of the system (27). The last gene of the metabolic operon, abfA, and the xsa gene most probably encode arabinofuranosidases involved in the intracellular degradation of arabinose oligomers (36). The abnA gene encodes an extracellular endo-arabinanase that degrades the arabinose homoglycan arabinan (16).The pathways of L-arabinose utilization in B. subtilis and Escherichia coli are identical, and the catabolic enzymes are functionally ...

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“…The BamHIEcoRI fragment containing the p petE -kgtP fusion was subcloned into the shuttle vector pRL25c. pRL25c, like pAM1691, is a derivative of pDU1; the copy number of pAM1691 is about 17 per chromosome (Lee et al, 2003). The construct was conjugated into Anabaena sp.…”

Section: Methodsmentioning

confidence: 99%

An increase in the level of 2-oxoglutarate promotes heterocyst development in the cyanobacterium Anabaena sp. strain PCC 7120

Laurent

Konde

et al. 2003

Microbiology

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In the filamentous cyanobacterium Anabaena sp. strain PCC 7120, a starvation of combined nitrogen induces differentiation of heterocysts, cells specialized in nitrogen fixation. How do filaments perceive the limitation of the source of combined nitrogen, and what determines the proportion of heterocysts? In cyanobacteria, 2-oxoglutarate provides a carbon skeleton for the incorporation of inorganic nitrogen. Recently, it has been proposed that the concentration of 2-oxoglutarate reflects the nitrogen status in cyanobacteria. To investigate the effect of 2-oxoglutarate on heterocyst development, a heterologous gene encoding a 2-oxoglutarate permease under the control of a regulated promoter was expressed in Anabaena sp. PCC 7120. The increase of 2-oxoglutarate within cells can trigger heterocyst differentiation in a subpopulation of filaments even in the presence of nitrate. In the absence of a source of combined nitrogen, it can increase heterocyst frequency, advance the timing of commitment to heterocyst development and further increase the proportion of heterocysts in a patS mutant. Here, it is proposed that the intracellular concentration of 2-oxoglutarate is involved in the determination of the proportion of the two cell types according to the carbon/nitrogen status of the filament.

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PlmA, a New Member of the GntR Family, Has Plasmid Maintenance Functions in Anabaena sp. Strain PCC 7120

Cited by 63 publications

References 54 publications

patS Minigenes Inhibit Heterocyst Development of Anabaena sp. Strain PCC 7120

patS Minigenes Inhibit Heterocyst Development of Anabaena sp. Strain PCC 7120

Functional Domains of theBacillus subtilisTranscription Factor AraR and Identification of Amino Acids Important for Nucleoprotein Complex Assembly and Effector Binding

An increase in the level of 2-oxoglutarate promotes heterocyst development in the cyanobacterium Anabaena sp. strain PCC 7120

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