Photosynthesis Research

, Volume 98, Issue 1–3, pp 337–347 | Cite as

Directed mutagenesis of the transmembrane domain of the PsbL subunit of photosystem II in Synechocystis sp. PCC 6803

Regular Paper

Abstract

The PsbL protein is one of three low-molecular-weight subunits identified at the monomer–monomer interface of photosystem II (PSII) [Ferreira et al. (2004) Science 303:1831–1838; Loll et al. (2005) Nature 438:1040–1044]. We have employed site-directed mutagenesis to investigate the role of PsbL in Synechocystis sp. PCC 6803 cells. Truncation of the C-terminus by deleting the last four residues (Tyr-Phe-Phe-Asn) prevented association of PsbL with the CP43-less monomeric sub-complex and therefore blocked PSII assembly resulting in an obligate photoheterotrophic strain. Replacement of these residues with Ala created four photoautotrophic mutants. Compared to wild type, the F37A, F38A, and N39A strains had reduced levels of assembled PSII centers and F37A and F38A cells were readily photodamaged. In contrast, Y36A and Y36F mutants were similar to wild type. However, each of these strains had elevated levels of the CP43-less inactive monomeric complex. Mutations targeting a putative hydrogen bond between Arg-16 and sulfoquinovosyldiacylglycerol resulted in mutants that were also highly susceptible to photodamage. Similarly mutations targeting a conserved Tyr residue (Tyr-20) also destabilized PSII under high light and suggest that Tyr-20–lipid interactions or interactions of Tyr-20 with PsbT influence the ability of PSII to recover from photodamage.

Keywords

Photosystem II Photodamage PsbL Site-directed mutagenesis Synechocystis sp. PCC 6803 

Abbreviations

BN-PAGE

Blue-native polyacrylamide gel electrophoresis

DCMU

3-(3,4-dichlorophenyl)-1,1-dimethylurea

DM

n-dodecyl β-d-maltoside

DMBQ

2,5-dimethyl-p-benzoquinone

EDTA

Ethylenediamine tetra-acetic acid (di-sodium salt)

HEPES

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

Kb

Kilobases

kDa

Kilodaltons

OD

Optical density

PCC

Pasteur Culture Collection

PCR

Polymerase chain reaction

PQ

Plastoquinone

PSII

Photosystem II

QA

Primary quinone electron acceptor of photosystem II

SDS

Sodium dodecyl sulfate

SDS-PAGE

SDS polyacrylamide gel electrophoresis

TES

2-[tris(hydroxymethyl)methyl]amino-1-ethanesulfonic acid

Tris

Tris(hydroxymethyl)aminomethane

Notes

Acknowledgment

This work was supported by a grant (UOO309) from the New Zealand Marsden Fund to J.J.E.-R.

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.Department of BiochemistryUniversity of OtagoDunedinNew Zealand

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