Abstract
High-light-inducible proteins (Hlips) are single-helix transmembrane proteins that are essential for the survival of cyanobacteria under stress conditions. The model cyanobacterium Synechocystis sp. PCC 6803 contains four Hlip isoforms (HliA-D) that associate with Photosystem II (PSII) during its assembly. HliC and HliD are known to form pigmented (hetero)dimers that associate with the newly synthesized PSII reaction center protein D1 in a configuration that allows thermal dissipation of excitation energy. Thus, it is expected that they photoprotect the early steps of PSII biogenesis. HliA and HliB, on the other hand, bind the PSII inner antenna protein CP47, but the mode of interaction and pigment binding have not been resolved. Here, we isolated His-tagged HliA and HliB from Synechocystis and show that these two very similar Hlips do not interact with each other as anticipated, rather they form HliAC and HliBC heterodimers. Both dimers bind Chl and β-carotene in a quenching conformation and associate with the CP47 assembly module as well as later PSII assembly intermediates containing CP47. In the absence of HliC, the cellular levels of HliA and HliB were reduced, and both bound atypically to HliD. We postulate a model in which HliAC-, HliBC-, and HliDC-dimers are the functional Hlip units in Synechocystis. The smallest Hlip, HliC, acts as a ‘generalist’ that prevents unspecific dimerization of PSII assembly intermediates, while the N-termini of ‘specialists’ (HliA, B or D) dictate interactions with proteins other than Hlips.
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The raw MS data has been submitted to the MassIVE repository (CCMS, University of California, San Diego) and can be accessed via the link ftp://massive.ucsd.edu/MSV000088753/.
Change history
23 March 2023
A Correction to this paper has been published: https://doi.org/10.1007/s11120-023-01016-y
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Acknowledgements
European Research Council Synergy Award 854126 and the Czech Science Foundation, grant No. 19-29225X funded this work. RS also acknowledges institutional support RVO 61388971. PK acknowledges support from the Czech Ministry of Education, Youth and Sports project ‘Mechanisms and dynamics of macromolecular complexes: from single molecules to cells’, CZ.02.1.01/0.0/0.0/15_003/0000441. Jan Pilný is thanked for HPLC analysis of pigments.
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MMK and RS designed the study; AW, MMK, and PK performed the experiments; MMK, PK, and RS analyzed the data; MMK and RS wrote the paper. All authors read and accepted the final manuscript.
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Konert, M.M., Wysocka, A., Koník, P. et al. High-light-inducible proteins HliA and HliB: pigment binding and protein–protein interactions. Photosynth Res 152, 317–332 (2022). https://doi.org/10.1007/s11120-022-00904-z
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DOI: https://doi.org/10.1007/s11120-022-00904-z