Chloroplast protease/chaperone AtDeg2 influences cotyledons opening and reproductive development in Arabidopsis

AtDeg2 is a chloroplast protein with dual protease/chaperone activity. Since data on how the individual activities of AtDeg2 affect growth and development of Arabidopsis thaliana plants is missing, two transgenic lines were prepared that express mutated AtDeg2 versions that have either only protease or chaperone activity and a comprehensive ontogenesis stage-based study was performed comprising wild type (WT) plants and insertional mutants that do not express AtDeg2, as well as the two transgenic lines. The repression of both AtDeg2 activities in deg2-3 mutants altered just a few phenotypic traits including the time when cotyledons were fully opened, the time when 10% flowers were open as well as the number of inflorescence branches and seed length in plants which have completed their generative development. It was demonstrated that complete opening of cotyledons as well as the number of inflorescence branches and seed length in plants which have completed their generative development required involvement of both AtDeg2 activities, whereas the time when 10% of flowers were open was controlled by AtDeg2 protease activity. These results show for the first time that the chaperone activity of AtDeg2 is needed for some elements of generative development of A. thaliana plants to proceed normally. So far, the chaperone activity of AtDeg2 was confirmed based on in vitro assays only

The structure, functions and degradation of pigment-binding proteins of photosystem II

Eleven proteins belonging to photosystem II (PSII) bind photosynthetic pigments in the form of thylakoid membrane-associated pigment-protein complexes. Five of them (PsbA, PsbB, PsbC, PsbD and PsbS) are assigned to PSII core complex while the remaining six (Lhcb1, Lhcb2, Lhcb3, Lhcb4, Lhcb5 and Lhcb6) constitute, along with their pigments, functional complexes situated more distantly with regard to P680 - the photochemical center of PSII. The main function of the pigment-binding proteins is to harvest solar energy and deliver it, in the form of excitation energy, ultimately to P680 although individual pigment-proteins may be engaged in other photosynthesis-related processes as well. The aim of this review is to present the current state of knowledge regarding the structure, functions and degradation of this family of proteins

Nitrate reduction and nitrogen fixation in symbiotic association Rhizobium - legumes.

The inhibitory effect of nitrate on nitrogenase activity in root nodules of legume plants has been known for a long time. The major factor inducing changes in nitrogenase activity is the concentration of free oxygen inside nodules. Oxygen availability in the infected zone of nodule is limited, among others, by the gas diffusion resistance in nodule cortex. The presence of nitrate may cause changes in the resistance to O2 diffusion. The aim of this paper is to review literature data concerning the effect of nitrate on the symbiotic association between rhizobia and legume plants, with special emphasis on nitrogenase activity. Recent advances indicate that symbiotic associations of Rhizobium strains characterized by a high nitrate reductase activity are less susceptible to inhibition by nitrate. A thesis may be put forward that dissimilatory nitrate reduction, catalyzed by bacteroid nitrate reductase, significantly facilitates the symbiotic function of bacteroids

S2P2—the chloroplast-located intramembrane protease and its impact on the stoichiometry and functioning of the photosynthetic apparatus of A. thaliana

S2P2 is a nuclear-encoded protease, potentially located in chloroplasts, which belongs to the zinc-containing, intramembrane, site-2 protease (S2P) family. In A. thaliana cells, most of the S2P proteases are located within the chloroplasts, where they play an important role in the development of chloroplasts, maintaining proper stoichiometric relations between polypeptides building photosynthetic complexes and influencing the sensitivity of plants to photoinhibitory conditions. Among the known chloroplast S2P proteases, S2P2 protease is one of the least known. Its exact location within the chloroplast is not known, nor is anything known about its possible physiological functions. Therefore, we decided to investigate an intra-chloroplast localization and the possible physiological role of S2P2. To study the intra-chloroplast localization of S2P2, we used specific anti-S2P2 antibodies and highly purified chloroplast fractions containing envelope, stroma, and thylakoid proteins. To study the physiological role of the protease, we used two lines of insertion mutants lacking the S2P2 protease protein. Here, we present results demonstrating the thylakoid localization of S2P2. Moreover, we present experimental evidence indicating that the lack of S2P2 in A. thaliana chloroplasts leads to a significant decrease in the level of photosystem I and photosystem II core proteins: PsaB, PsbA, PsbD, and PsbC, as well as polypeptides building both the main light-harvesting antenna (LHC II), Lhcb1 and Lhcb2, as well as Lhcb4 and Lhcb5 polypeptides, constituting elements of the minor, peripheral antenna system. These changes are associated with a decrease in the number of PS II–LHC II supercomplexes. The consequence of these disorders is a greater sensitivity of s2p2 mutants to photoinhibition. The obtained results clearly indicate that the S2P2 protease is another thylakoid protein that plays an important role in the proper functioning of A. thaliana chloroplasts, especially in high-light-intensity conditions

The contribution of individual domains of chloroplast protein AtDeg2 to its chaperone and proteolytic activities

The thylakoid protease AtDeg2 is a non-ATP hydrolyzing chloroplast protease/chaperone peripherally connected with stromal side of thylakoid membrane. Its linear structure consists of protease domain and two PDZ domains. To unveil the significance of individual domains, chaperone and proteolytic activities of AtDeg2, its mutated recombinant versions have been developed and their ability to suppress protein aggregation and resolubilization of protein aggregates as well as the ability to degrade substrate protein was examined in vitro. Our work reveals for the first time that AtDeg2 is able not only to suppress aggregation of denatured proteins, but to resolubilize existing protein aggregates as well. We show that PDZ2 domain contributes significantly to both chaperone and protease activities of AtDeg2, whereas PDZ1 is required for chaperone but superfluous for proteolytic activity. Protease domain – but not S-268 in its catalytic site – contributes to chaperone activities of AtDeg2. These results show an entirely new function of AtDeg2 chaperone/protease (i.e., disaggregation of protein aggregates) and allow to identify structural motifs required for “old” and new functions of AtDeg2