Roles of PsbI and PsbM in photosystem II dimer formation and stability studied by deletion mutagenesis and X-ray crystallography

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Abstract

PsbM and PsbI are two low molecular weight subunits of photosystem II (PSII), with PsbM being located in the center, and PsbI in the periphery, of the PSII dimer. In order to study the functions of these two subunits from a structural point of view, we crystallized and analyzed the crystal structure of PSII dimers from two mutants lacking either PsbM or PsbI. Our results confirmed the location of these two subunits in the current crystal structure, as well as their absence in the respective mutants. The relative contents of PSII dimers were found to be decreased in both mutants, with a concomitant increase in the amount of PSII monomers, suggesting a destabilization of PSII dimers in both of the mutants. On the other hand, the accumulation level of the overall PSII complexes in the two mutants was similar to that in the wild-type strain. Treatment of purified PSII dimers with lauryldimethylamine N-oxide at an elevated temperature preferentially disintegrated the dimers from the PsbM deletion mutant into monomers and CP43-less monomers, whereas no significant degradation of the dimers was observed from the PsbI deletion mutant. These results indicate that although both PsbM and PsbI are required for the efficient formation and stability of PSII dimers in vivo, they have different roles, namely, PsbM is required directly for the formation of dimers and its absence led to the instability of the dimers accumulated. On the other hand, PsbI is required in the assembly process of PSII dimers in vivo; once the dimers are formed, PsbI was no longer required for its stability.

Research highlights

► Location of PsbM and PsbI in PSII was confirmed. ► Deletion of both PsbM and PsbI decreased the stability of PSII dimer. ► PsbM and PsbI played different roles in the formation and stability of PSII dimers.

Abbreviations

Chl
chlorophyll
DDM
n-dodecyl-β-D-maltoside
hrCNE
high resolution clear native electrophoresis
LDAO
lauryldimethylamine N-oxide
Mes
4-morpholineethanesulfonic acid
PSII
photosystem II
PSIIcc
photosystem II core complex
SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis

Keywords

Photosystem II
Mutant
Crystal structure
PsbM
PsbI
Oxygen evolution

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