Over-production of the D1 protein of photosystem II reaction centre in the cyanobacterium Synechococcus sp. PCC 7942.
The unicellular cyanobacterium Synechococcus sp. PCC 7942 has three psbA genes encoding two different forms of the photosystem II reaction centre protein D1 (D1:1 and D1:2). The level of expression of these psbA genes and the synthesis of D1:1 and D1:2 are strongly regulated under varying light conditions. In order to better understand the regulatory mechanisms underlying these processes, we have constructed a strain of Synechococcus sp. PCC 7942 capable of over-producing psbA mRNA and D1 protein. In this study, we describe the over-expression of D1:1 using a tac-hybrid promoter in front of the psbAI gene in combination with lacIQ repressor system. Over-production of D1:1 was induced by growing cells for 12 h at 50 mumol photons m-2 s-1 in the presence of 40 or 80 micrograms/ml IPTG. The amount of psbAI mRNA and that of D1:1 protein in cells grown with IPTG was three times and two times higher, respectively. A higher concentration of IPTG (i.e., 150 micrograms/ml) did not further increase the production of the psbAI message or D1:1. The over-production of D1:1 caused a decrease in the level of D1:2 synthesised, resulting in most PSII reaction centres containing D1:1. However, the over-production of D1:1 had no effect on the pigment composition (chlorophyll a or phycocyanin/number of cells) or the light-saturated rate of photosynthesis. This and the fact that the total amounts of D1 and D2 proteins were not affected by IPTG suggest that the number of PSII centres within the membranes remained unchanged. From these results, we conclude that expression of psbAI can be regulated by using the tac promoter and lacIQ system. However, the accumulation of D1:1 protein into the membrane is regulated by the number of PSII centres.
Soitamo
A J
AJ
Department of Biology, University of Turku, Finland.
Zhou
G
G
Clarke
A K
AK
Oquist
G
G
Aro
E M
EM
Gustafsson
P
P
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002462:Cell Membrane / Q000378:metabolism; D000458:Cyanobacteria / Q000378:metabolism*; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D018506:Gene Expression Regulation, Plant; D015151:Immunoblotting; D007700:Kinetics; D045342:Light-Harvesting Protein Complexes; D008969:Molecular Sequence Data; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000096:biosynthesis*; D045332:Photosystem II Protein Complex; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D015183:Restriction Mapping
D001426:Bacterial Proteins; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D012333:RNA, Messenger