The PsbL, PsbM and PsbT subunits of photosystem II (PSII) are single-helix membrane-spanning proteins found at the monomer-monomer interface that may stabilize the dimeric complex. This study has characterised strains of Synechocystis sp. PCC 6803 where psbL, psbM and psbT have been interrupted by the insertion of antibiotic-resistance cassettes. The [Delta]PsbL strain exhibited slowed growth that correlated with a disruption in PSII assembly leading to an accumulation of CP43-less PSII monomers. Moreover, the [Delta]PsbL:[Delta]PsbM and [Delta]PsbL:[Delta]PsbT double mutants were not photoautotrophic. In contrast, the [Delta]PsbM and [Delta]PsbT strains grew photoautotrophically and supported oxygen evolution, albeit at reduced rates compared to wild type. S-state analyses showed that the removal of PsbM or PsbT did not affect the donor side reactions of PSII, which includes the oxidation of water, however, the removal of PsbT impaired electron flow between Q[A] and Q[B] on the acceptor side of PSII. Blue-Native PAGE revealed that removal of either PsbM or PsbT was insufficient to entirely disrupt dimer formation; however, the combined removal of PsbM and PsbT resulted in the predominance of monomeric forms of PSII in the [Delta]PsbM:[Delta]PsbT strain. Under high light (2 mE m⁻� s⁻� at 30�C), [Delta]PsbM and [Delta]PsbT cells were considerably more susceptible to photoinactivation than wild type; however, they were able to fully recover in a protein synthesis-dependent manner when returned to moderate light levels (0.03 mE m⁻� s⁻�). A requirement for Psb27 was found in the protein-synthesis-dependent recovery of photoinactivated [Delta]PsbT cells. More significantly, an absolute functional requirement was found for Psb27 in the [Delta]PsbM strain, where functional PSII complexes are not assembled in the absence of Psb27. These results suggest that Psb27 is critical for PSII assembly in the absence of PsbM, and also for the protein-synthesis-dependent recovery of PSII in the absence of PsbT. Moreover, in addition to Psb27, the PsbU subunit of the oxygen-evolving complex was also found to be an absolute functional requirement in the [Delta]PsbM strain, where functional PSII centres are not assembled when both PsbM and PsbU are absent. It appears, therefore, that PsbM has crucial functional interactions with specific extrinsic proteins located in the vicinity of the oxygen-evolving complex. Interestingly, the [Delta]PsbM strain was also found to have a high susceptibility to suppressor mutations, indicating it has important functional roles in the cyanobacterial cell.
| Identifer | oai:union.ndltd.org:ADTP/266364 |
| Date | January 2008 |
| Creators | Bentley, Fiona K, n/a |
| Publisher | University of Otago. Department of Biochemistry |
| Source Sets | Australiasian Digital Theses Program |
| Language | English |
| Detected Language | English |
| Rights | http://policy01.otago.ac.nz/policies/FMPro?-db=policies.fm&-format=viewpolicy.html&-lay=viewpolicy&-sortfield=Title&Type=Academic&-recid=33025&-find), Copyright Fiona K Bentley |
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