Why have organelles retained genomes?

The observation that chloroplasts and mitochondria have retained relics of eubacterial genomes and a protein-synthesizing machinery has long puzzled biologists. If most genes have been transferred from organelles to the nucleus during evolution, why not all? What selective pressure maintains genomes in organelles? Electron transport through the photosynthetic and respiratory membranes is a powerful - but dangerous - source of energy. Recent evidence suggests that organelle genomes have persisted because structural proteins that maintain redox balance within bioenergetic membranes must be synthesized when and where they are needed, to counteract the potentially deadly side effects of ATP-generating electron transport.

A protein tyrosine kinase of chloroplast thylakoid membranes phosphorylates light harvesting complex II proteins.

Phosphorylation of chloroplast thylakoid proteins, in particular light harvesting complex II (LHC II), is believed to play an important role in regulating photosynthetic electron transfer. Evidence supporting the involvement of multiple protein kinases in this system is mounting. We have re-examined pea thylakoid membranes and found evidence for a membrane-associated protein tyrosine kinase (PTK). Phosphorylation of many thylakoid proteins, including LHC II, is sensitive to treatment with the tyrosine kinase inhibitor genistein. Anti-phosphotyrosine antibodies react specifically with nine thylakoid proteins, two of which have been identified as components of LHC II. The phosphate associated with these two proteins is also resistant to strong base and acid treatment, further substantiating the assignment of phosphotyrosine. Potential interactions between this novel chloroplast PTK activity and the well-documented threonine kinase activities are discussed and the presence of a cascade of thylakoid protein kinases is proposed.

Truncated recombinant light harvesting complex II proteins are substrates for a protein kinase associated with photosystem II core complexes.

Previous studies directed towards understanding phosphorylation of the chlorophyll alb binding proteins comprising light harvesting complex II (LHC II) have concentrated on a single phosphorylation site located close to the N-terminus of the mature proteins. Here we show that a series of recombinant pea Lhcb1 proteins, each missing an N-terminal segment including this site, are nevertheless phosphorylated by a protein kinase associated with a photosystem II core complex preparation. An Lhch1 protein missing the first 58 amino acid residues is not, however, phosphorylated. The results demonstrate that the LHC II proteins are phosphorylated at one or more sites, the implications of which are discussed.

Minor constituents of photosystem II core complexes: possible regulators of photosystem II protein kinase.

We recently identified a 58-kDa protein kinase, PS II-PK, closely associated in substoichiometric abundance with a core complex of photosystem II and capable of phosphorylating both the photosystem and its associated light-harvesting proteins. Oxidizing species, produced during aerobic illumination, inhibited the kinase, and the irreversibility of this process is now demonstrated. Other substoichiometric protein constituents of the core preparation were identified as probably originating in the grana margins. These include the cytochrome b6/f complex, the apocytochrome f precursor, and membrane-bound ferredoxin:NADP+ oxidoreductase. PS II-PK was successfully resolved from photosystem II cores and shown to be active in the absence of cytochrome complex.

A protein kinase in the core of photosystem II.

In green plants, several intrinsic protein components of the photosystem II (PS II) complexes are subject to reversible phosphorylation on threonine residues. Evidence from mutant and inhibitor studies indicates that multiple kinases are involved. The protein kinases appear to be membrane-bound and redox-regulated, with activity requiring reducing conditions. We report the identification of a protein kinase activity which copurifies with a core complex of PS II and is capable of phosphorylating the photosystem proteins and associated light-harvesting complex. The enzyme is a distinct and novel protein whose close proximity to the photosystem reaction center is confirmed by its rapid inactivation under strong red light irradiation in the presence of oxygen.

A 64-kDa protein is a substrate for phosphorylation by a distinct thylakoid protein kinase.

Solubilization of spinach thylakoids with the nonionic detergent n-octyl-ß-D-glucopyranoside (OG) releases active protein kinase from the membrane. Further purification was reported to demonstrate that a 64-kDa protein is the origin of this kinase activity (Coughlan S J and Hind G (1986) J Biol Chem 261: 11378-11385). The N-terminal sequence of this protein was subsequently determined (Gal A, Herrmann R, Lottspiech F and Ohad I (1992) FEBS Lett 298: 33-35). Liquid phase isoelectric focusing of the OG extract and an hydroxylapatite-purified fraction, derived from the OG preparation, reveals that the 64-kDa protein with this documented N-terminal sequence can be separated from the protein kinase activity. Experimental conditions were optimised by manipulation of ampholyte and detergent concentrations to maximise protein solubility and enzyme activity. The kinase-containing fraction was able to catalyze the phosphorylation of several proteins including the 64-kDa which was identified using antibodies raised against a synthetic peptide corresponding to the N-terminal sequence. The results described indicate that this 64-kDa protein is not the protein kinase responsible for the phosphorylation of the light-harvesting complex associated with Photosystem II.

Identification of the psbH gene product as a 6 kDa phosphoprotein in the cyanobacterium Synechocystis 6803.

The product of the psbH gene has been identified in Synechocystis 6803 thylakoid membranes as a 6 kDa phosphoprotein. This protein becomes phosphorylated in vitro despite the fact that in cyanobacteria it is truncated at the N-terminus such that the phosphorylation site identified in the higher plant protein is missing. Phosphorylation occurred both in the light and in the dark but was inhibited by oxidising conditions, DCMU and zinc ions. The cyanobacterial 6 kDa phosphoprotein degrades when the membranes are subjected to high intensity illumination.

Chlorophyll levels in the pigment-binding proteins of photosystem II. A study based on the chlorophyll to cytochrome ratio in different photosystem II preparations.

The chlorophyll levels in pigment proteins of photosystem II were investigated by using photosystem II preparations with different levels of complexity. Based on the assumption that there is 1 cytochrome b559 per reaction centre it has been found that oxygen-evolving complexes containing CP26 and CP29 bind 42 chlorophyll molecules. When CP26 and CP29 are stripped away, the resulting PSII cores bind 30 chlorophyll molecules while CP43-less cores bind approximately 18 chlorophylls. It is therefore concluded that CP47 and CP43 bind 9-12 molecules of chlorophyll a and the D1/D2 complex binds 6 chlorophylls. Taken together CP26 and CP29 bind about 12 chlorophyll molecules.