Photosynthetic organisms are continuously subjected to changes in light quantity and quality, and must adjust their photosynthetic machinery so that it maintains optimal performance under limiting light and minimizes photodamage under excess light. To achieve this goal, these organisms use two main strategies in which light-harvesting complex II (LHCII), the light-harvesting system of photosystem II (PSII), plays a key role both for the collection of light energy and for photoprotection. The first is energy-dependent nonphotochemical quenching, whereby the high-light-induced proton gradient across the thylakoid membrane triggers a process in which excess excitation energy is harmlessly dissipated as heat. The second involves a redistribution of the mobile LHCII between the two photosystems in response to changes in the redox poise of the electron transport chain sensed through a signaling chain. These two processes strongly diminish the production of damaging reactive oxygen species, but photodamage of PSII is unavoidable, and it is repaired efficiently.

An inducible chloroplast gene expression system was developed in Chlamydomonas reinhardtii by taking advantage of the properties of the copper-sensitive cytochrome c ₆ promoter and of the nucleus-encoded Nac2 chloroplast protein. This protein is specifically required for the stable accumulation of the chloroplast psbD RNA and acts on its 5′ UTR. A construct containing the Nac2 coding sequence fused to the cytochrome c ₆ promoter was introduced into the nac2-26 mutant strain deficient in Nac2 . In this transformant, psbD is expressed in copper-depleted but not in copper-replete medium. Because psbD encodes the D2 reaction center polypeptide of photosystem II (PSII), the repression of psbD leads to the loss of PSII. We have tested this system for hydrogen production. Upon addition of copper to cells pregrown in copper-deficient medium, PSII levels declined to a level at which oxygen consumption by respiration exceeded oxygen evolution by PSII. The resulting anaerobic conditions led to the induction of hydrogenase activity. Because the Cyc6 promoter is also induced under anaerobic conditions, this system opens possibilities for sustained cycling hydrogen production. Moreover, this inducible gene expression system is applicable to any chloroplast gene by replacing its 5′ UTR with the psbD 5′ UTR in the same genetic background. To make these strains phototrophic, the 5′ UTR of the psbD gene was replaced by the petA 5′ UTR. As an example, we show that the reporter gene aadA driven by the psbD 5′ UTR confers resistance to spectinomycin in the absence of copper and sensitivity in its presence in the culture medium.

State transitions allow for the balancing of the light excitation energy between photosystem I and photosystem II and for optimal photosynthetic activity when photosynthetic organisms are subjected to changing light conditions. This process is regulated by the redox state of the plastoquinone pool through the Stt7/STN7 protein kinase required for phosphorylation of the light-harvesting complex LHCII and for the reversible displacement of the mobile LHCII between the photosystems. We show that Stt7 is associated with photosynthetic complexes including LHCII, photosystem I, and the cytochrome b₆f complex. Our data reveal that Stt7 acts in catalytic amounts. We also provide evidence that Stt7 contains a transmembrane region that separates its catalytic kinase domain on the stromal side from its N-terminal end in the thylakoid lumen with two conserved Cys that are critical for its activity and state transitions. On the basis of these data, we propose that the activity of Stt7 is regulated through its transmembrane domain and that a disulfide bond between the two lumen Cys is essential for its activity. The high-light–induced reduction of this bond may occur through a transthylakoid thiol–reducing pathway driven by the ferredoxin-thioredoxin system which is also required for cytochrome b₆f assembly and heme biogenesis. To grow optimally, photosynthetic organisms need to constantly adjust to changing light conditions. One of these adjustments, called state transitions, allows light energy to be redistributed between the two photosynthetic reaction center complexes in a cell's chloroplasts. These complexes act in concert with other components of the photosynthetic machinery to turn light energy into cellular energy. A key component in the regulation of state transitions is the chloroplast protein Stt7 (also known as STN7), which can modify other proteins by adding a phosphate group. When light levels change, the oxidation level of a pool of another chloroplast component, plastoquinone, changes, which in turn activates Stt7, inducing it to phosphorylate specific proteins of the light-harvesting complex of one reaction center. As a result, a portion of this light-harvesting complex is transferred from one photosynthetic reaction center to the other, thereby optimizing photosynthetic efficiency. Here, we have addressed the configuration of Stt7 within the thylakoid membrane of the chloroplast and the molecular mechanisms underlying its activation. Our data reveal that the level of Stt7 protein changes drastically under specific environmental conditions, that the protein does not need to be present in a one-to-one ratio with its targets for activity, and that it associates directly with a number of components of the photosynthetic machinery. The protein-modifying domain of Stt7 is exposed to the outer side of the thylakoid membrane, whereas the domain critical for regulation of its activity lies on the inner side of the thylakoid membrane. These results shed light on the molecular mechanisms that allow photosynthetic organisms to adjust to fluctuations in light levels.