Abstract
The ATP-binding cassette (ABC) transporter is a multi-subunit membrane protein complex involved in lipid transport and acid stress tolerance in the cyanobacterium Synechocystis sp. PCC 6803. This organism has two sets of three ABC transporter subunits: Slr1045 and Slr1344, Sll0751 and Sll1002, and Sll1001 and Sll1041. We previously found that Slr1045 is essential for survival under acid stress condition (Tahara et al. 2012). In the present study, we examined the participation of other ABC transporter subunits in acid stress tolerance using a deletion mutant series of Synechocystis sp. PCC 6803. Although Slr1344 is highly homologous to Slr1045, Δslr1344 cells were not susceptible to acid stress. Δsll0751 and Δsll1041 cells displayed acid stress sensitivity, whereas Δsll1001/sll1002 double mutant cells grew normally. Under high- and low-temperature stress conditions, the growth rate of Δslr1344 and Δsll1001/sll1002 cells did not differ from WT cells, whereas Δsll0751 and Δsll1041 cells showed significant growth retardation, as previously observed in Δslr1045 cells. Moreover, nile red staining showed more lipid accumulation in Δslr1045, Δsll0751, and Δsll1041 cells than in WT cells. These results suggest that Slr1045, Sll0751, and Sll1041 function together as a lipid transport complex in Synechocystis sp. PCC 6803 and are essential for growth under various stresses.
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Abbreviations
- ABC:
-
ATP-binding cassette
- PG:
-
Phosphatidylglycerol
- PM:
-
Plasma membrane
- Synechocystis :
-
Synechocystis sp. PCC 6803
- TM:
-
Thylakoid membrane
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This study was supported by the Program for Development of Strategic Research Center in Private Universities, supported by MEXT (S1001020).
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11120_2015_153_MOESM1_ESM.tif
Multiple sequence alignment of Slr1045, Slr1344, MlaE, and TGD1 protein. Conserved amino acid sequences are shown by black boxes. Asterisks indicate amino acid sequence pairs with the same sequence. Slr1045 and Slr1344 are more similar to TGD1 than to MlaE. Slr1045 has homology to MlaE (38 % identity and 57 % positive), which plays a role in the maintenance of outer membrane lipid asymmetry in E. coli, and TGD1 (53 % identity and 73 % positive), which is a phosphatidic acid transporter in A. thaliana. Slr1344 protein has homology to MlaE (32 % identity and 54 % positive) and TGD1 (50 % identity and 71 % positive). These proteins may have the same function (TIFF 85227 kb)
11120_2015_153_MOESM2_ESM.tif
Construction of slr1344, sll1001/sll1002, sll0751, and sll1041 mutant cells. PCR analyses using chromosomal DNA template for WT and mutant genes: slr1344 (a), sll1002 and sll1001 (b), sll0751 (c), and sll1041 (d). Primers specific for each gene were used to confirm chromosomal DNA disruption in mutants (TIFF 40575 kb)
11120_2015_153_MOESM3_ESM.tif
Histogram of observed bright spots. WT, Δslr1045, Δslr1344, Δsll1001/sll1002, Δsll0751, and Δsll1041 cells were cultivated on BG-11 medium (pH 8.0) for 7 days at 30 °C. Stained cells were then observed by fluorescence microscopy with optical filters. The number of bright spots in cells was counted. (TIFF 20078 kb)
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Tahara, H., Matsuhashi, A., Uchiyama, J. et al. Sll0751 and Sll1041 are involved in acid stress tolerance in Synechocystis sp. PCC 6803. Photosynth Res 125, 233–242 (2015). https://doi.org/10.1007/s11120-015-0153-6
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DOI: https://doi.org/10.1007/s11120-015-0153-6