Dataset Information

Identification of an ATP-binding cassette transporter involved in bicarbonate uptake in the cyanobacterium Synechococcus sp. strain PCC 7942.

ABSTRACT: Exposure of cells of cyanobacteria (blue-green algae) grown under high-CO(2) conditions to inorganic C-limitation induces transcription of particular genes and expression of high-affinity CO(2) and HCO(3)(-) transport systems. Among the low-CO(2)-inducible transcription units of Synechococcus sp. strain PCC 7942 is the cmpABCD operon, encoding an ATP-binding cassette transporter similar to the nitrate/nitrite transporter of the same cyanobacterium. A nitrogen-regulated promoter was used to selectively induce expression of the cmpABCD genes by growth of transgenic cells on nitrate under high CO(2) conditions. Measurements of the initial rate of HCO(3)(-) uptake after onset of light, and of the steady-state rate of HCO(3)(-) uptake in the light, showed that the controlled induction of the cmp genes resulted in selective expression of high-affinity HCO(3)(-) transport activity. The forced expression of cmpABCD did not significantly increase the CO(2) uptake capabilities of the cells. These findings demonstrated that the cmpABCD genes encode a high-affinity HCO(3)(-) transporter. A deletion mutant of cmpAB (M42) retained low CO(2)-inducible activity of HCO(3)(-) transport, indicating the occurrence of HCO(3)(-) transporter(s) distinct from the one encoded by cmpABCD. HCO(3)(-) uptake by low-CO(2)-induced M42 cells showed lower affinity for external HCO(3)(-) than for wild-type cells under the same conditions, showing that the HCO(3)(-) transporter encoded by cmpABCD has the highest affinity for HCO(3)(-) among the HCO(3)(-) transporters present in the cyanobacterium. This appears to be the first unambiguous identification and description of a primary active HCO(3)(-) transporter.

SUBMITTER: Omata T

PROVIDER: S-EPMC23989 | biostudies-literature | 1999 Nov

REPOSITORIES: biostudies-literature

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Publications

Identification of an ATP-binding cassette transporter involved in bicarbonate uptake in the cyanobacterium Synechococcus sp. strain PCC 7942.

Omata T T Price G D GD Badger M R MR Okamura M M Gohta S S Ogawa T T

Proceedings of the National Academy of Sciences of the United States of America 19991101 23

Exposure of cells of cyanobacteria (blue-green algae) grown under high-CO(2) conditions to inorganic C-limitation induces transcription of particular genes and expression of high-affinity CO(2) and HCO(3)(-) transport systems. Among the low-CO(2)-inducible transcription units of Synechococcus sp. strain PCC 7942 is the cmpABCD operon, encoding an ATP-binding cassette transporter similar to the nitrate/nitrite transporter of the same cyanobacterium. A nitrogen-regulated promoter was used to selec ...[more]

PMID: 10557362

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Project description:The heat shock protein CIpB (HSP100) is a member of the diverse group of Clp polypeptides that function as molecular chaperones and/or regulators of energy-dependent proteolysis. A single-copy gene coding for a ClpB homolog was cloned and sequenced from the unicellular cyanobacterium Synechococcus sp. strain PCC 7942. The predicted polypeptide sequence was most similar to sequences of cytosolic ClpB from bacteria and higher plants (i.e., 70 to 75%). Inactivation of clpB in Synechococcus sp. strain PCC 7942 resulted in no significant differences from the wild-type phenotype under optimal growth conditions. In the wild type, two forms of ClpB were induced during temperature shifts from 37 to 47.5 or 50 degrees C, one of 92 kDa, which matched the predicted size, and another smaller protein of 78 kDa. Both proteins were absent in the delta clpB strain. The level of induction of the two ClpB forms in the wild type increased with increasingly higher temperatures, while the level of the constitutive ClpC protein remained unchanged. In the delta clpB strain, however, the ClpC content almost doubled during the heating period, presumably to compensate for the loss of ClpB activity. Photosynthetic measurements at 47.5 and 50 degrees C showed that the null mutant was no more susceptible to thermal inactivation than the wild type. Using photosynthesis as a metabolic indicator, an assay was developed for Synechococcus spp. to determine the importance of ClpB for acquired thermotolerance. Complete inactivation of photosynthetic oxygen evolution occurred in both the wild type and the delta clpB strain when they were shifted from 37 directly to 55 degrees C for 10 min. By preexposing the cells at 50 degrees C for 1.5 h, however, a significant level of photosynthesis was retained in the wild type but not in the mutant after the treatment at 55 degrees C for 10 min. Cell survival determinations confirmed that the loss of ClpB synthesis caused a fivefold reduction in the ability of Synechococcus cells to develop thermotolerance. These results clearly show that induction of ClpB at high temperatures is vital for sustained thermotolerance in Synechococcus spp., the first such example for either a photosynthetic or a prokaryotic organism.

Dataset Information

Identification of an ATP-binding cassette transporter involved in bicarbonate uptake in the cyanobacterium Synechococcus sp. strain PCC 7942.

Publications

Identification of an ATP-binding cassette transporter involved in bicarbonate uptake in the cyanobacterium Synechococcus sp. strain PCC 7942.

Similar Datasets

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