An independent role of cytochrome c-550 in cyanobacterial photosystem II as revealed by double-deletion mutagenesis of the psbO and psbV genes in Synechocystis sp. PCC 6803

Jian-Ren Shen, Robert L. Burnap, Yorinao Inoue

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Cytochrome (cyt) c-550 and the 33 kDa protein are two extrinsic components that function in maintaining oxygen evolution in cyanobacterial cells. Deletion of either of the two components has been shown to result in cyanobacterial phenotypes that are still capable of photoautotrophic growth albeit with a reduced rate. In order to study the function of cyt c-550 in cyanobacterial photosystem II (PSII) and its possible interaction with the 33 kDa extrinsic protein, we constructed a mutant lacking both cyt c-550 and the 33 kDa protein by inactivating the psbV and psbO genes simultaneously in a cyanobacterium, Synechocystis sp. PCC 6803. The resultant double-deletion mutant was unable to grow photoautotrophically and showed almost no oxygen-evolving activity (less than 10% of the wild type). This residual activity was also lost rapidly upon illumination, suggesting an increased sensitivity of the mutant cells toward photoinhibition. Thermoluminescence measurements indicated that the mutant virtually cannot undergo normal charge accumulation (S-state transitions) leading to oxygen evolution. Herbicide-binding and Western blot analyses showed that the mutant accumulates the PSII complex to an extent of only 20% of that in wild-type cells. Combined with previous results, the present results indicated that cyt c-550 supported oxygen evolution in the single-deletion mutant lacking the 33 kDa protein alone and vice versa. Thus, both cyt c-550 and the 33 kDa protein function independently in maintaining cyanobacterial oxygen-evolving activity in vivo, and both of them are required for the optimal activity. This not only provides another conclusive line of evidence for the concept that cyt c-550 functions in cyanobacterial PSII oxygen evolution but also reveals a functional difference between this extrinsic cyt in cyanobacterial PSII and the extrinsic 23 kDa protein in higher plant PSII, as the 23 kDa protein alone cannot support oxygen evolution in the absence of the 33 kDa protein.

Original languageEnglish
Pages (from-to)12661-12668
Number of pages8
JournalBiochemistry
Volume34
Issue number39
Publication statusPublished - 1995
Externally publishedYes

Fingerprint

Synechocystis
Mutagenesis
Photosystem II Protein Complex
Cytochromes c
Genes
Oxygen
Proteins
Thermoluminescence
Cyanobacteria
Herbicides
Cytochromes
Lighting
Western Blotting
Phenotype

ASJC Scopus subject areas

  • Biochemistry

Cite this

An independent role of cytochrome c-550 in cyanobacterial photosystem II as revealed by double-deletion mutagenesis of the psbO and psbV genes in Synechocystis sp. PCC 6803. / Shen, Jian-Ren; Burnap, Robert L.; Inoue, Yorinao.

In: Biochemistry, Vol. 34, No. 39, 1995, p. 12661-12668.

Research output: Contribution to journalArticle

@article{aea15706911340b3bc919a57c229abf5,
title = "An independent role of cytochrome c-550 in cyanobacterial photosystem II as revealed by double-deletion mutagenesis of the psbO and psbV genes in Synechocystis sp. PCC 6803",
abstract = "Cytochrome (cyt) c-550 and the 33 kDa protein are two extrinsic components that function in maintaining oxygen evolution in cyanobacterial cells. Deletion of either of the two components has been shown to result in cyanobacterial phenotypes that are still capable of photoautotrophic growth albeit with a reduced rate. In order to study the function of cyt c-550 in cyanobacterial photosystem II (PSII) and its possible interaction with the 33 kDa extrinsic protein, we constructed a mutant lacking both cyt c-550 and the 33 kDa protein by inactivating the psbV and psbO genes simultaneously in a cyanobacterium, Synechocystis sp. PCC 6803. The resultant double-deletion mutant was unable to grow photoautotrophically and showed almost no oxygen-evolving activity (less than 10{\%} of the wild type). This residual activity was also lost rapidly upon illumination, suggesting an increased sensitivity of the mutant cells toward photoinhibition. Thermoluminescence measurements indicated that the mutant virtually cannot undergo normal charge accumulation (S-state transitions) leading to oxygen evolution. Herbicide-binding and Western blot analyses showed that the mutant accumulates the PSII complex to an extent of only 20{\%} of that in wild-type cells. Combined with previous results, the present results indicated that cyt c-550 supported oxygen evolution in the single-deletion mutant lacking the 33 kDa protein alone and vice versa. Thus, both cyt c-550 and the 33 kDa protein function independently in maintaining cyanobacterial oxygen-evolving activity in vivo, and both of them are required for the optimal activity. This not only provides another conclusive line of evidence for the concept that cyt c-550 functions in cyanobacterial PSII oxygen evolution but also reveals a functional difference between this extrinsic cyt in cyanobacterial PSII and the extrinsic 23 kDa protein in higher plant PSII, as the 23 kDa protein alone cannot support oxygen evolution in the absence of the 33 kDa protein.",
author = "Jian-Ren Shen and Burnap, {Robert L.} and Yorinao Inoue",
year = "1995",
language = "English",
volume = "34",
pages = "12661--12668",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "39",

}

TY - JOUR

T1 - An independent role of cytochrome c-550 in cyanobacterial photosystem II as revealed by double-deletion mutagenesis of the psbO and psbV genes in Synechocystis sp. PCC 6803

AU - Shen, Jian-Ren

AU - Burnap, Robert L.

AU - Inoue, Yorinao

PY - 1995

Y1 - 1995

N2 - Cytochrome (cyt) c-550 and the 33 kDa protein are two extrinsic components that function in maintaining oxygen evolution in cyanobacterial cells. Deletion of either of the two components has been shown to result in cyanobacterial phenotypes that are still capable of photoautotrophic growth albeit with a reduced rate. In order to study the function of cyt c-550 in cyanobacterial photosystem II (PSII) and its possible interaction with the 33 kDa extrinsic protein, we constructed a mutant lacking both cyt c-550 and the 33 kDa protein by inactivating the psbV and psbO genes simultaneously in a cyanobacterium, Synechocystis sp. PCC 6803. The resultant double-deletion mutant was unable to grow photoautotrophically and showed almost no oxygen-evolving activity (less than 10% of the wild type). This residual activity was also lost rapidly upon illumination, suggesting an increased sensitivity of the mutant cells toward photoinhibition. Thermoluminescence measurements indicated that the mutant virtually cannot undergo normal charge accumulation (S-state transitions) leading to oxygen evolution. Herbicide-binding and Western blot analyses showed that the mutant accumulates the PSII complex to an extent of only 20% of that in wild-type cells. Combined with previous results, the present results indicated that cyt c-550 supported oxygen evolution in the single-deletion mutant lacking the 33 kDa protein alone and vice versa. Thus, both cyt c-550 and the 33 kDa protein function independently in maintaining cyanobacterial oxygen-evolving activity in vivo, and both of them are required for the optimal activity. This not only provides another conclusive line of evidence for the concept that cyt c-550 functions in cyanobacterial PSII oxygen evolution but also reveals a functional difference between this extrinsic cyt in cyanobacterial PSII and the extrinsic 23 kDa protein in higher plant PSII, as the 23 kDa protein alone cannot support oxygen evolution in the absence of the 33 kDa protein.

AB - Cytochrome (cyt) c-550 and the 33 kDa protein are two extrinsic components that function in maintaining oxygen evolution in cyanobacterial cells. Deletion of either of the two components has been shown to result in cyanobacterial phenotypes that are still capable of photoautotrophic growth albeit with a reduced rate. In order to study the function of cyt c-550 in cyanobacterial photosystem II (PSII) and its possible interaction with the 33 kDa extrinsic protein, we constructed a mutant lacking both cyt c-550 and the 33 kDa protein by inactivating the psbV and psbO genes simultaneously in a cyanobacterium, Synechocystis sp. PCC 6803. The resultant double-deletion mutant was unable to grow photoautotrophically and showed almost no oxygen-evolving activity (less than 10% of the wild type). This residual activity was also lost rapidly upon illumination, suggesting an increased sensitivity of the mutant cells toward photoinhibition. Thermoluminescence measurements indicated that the mutant virtually cannot undergo normal charge accumulation (S-state transitions) leading to oxygen evolution. Herbicide-binding and Western blot analyses showed that the mutant accumulates the PSII complex to an extent of only 20% of that in wild-type cells. Combined with previous results, the present results indicated that cyt c-550 supported oxygen evolution in the single-deletion mutant lacking the 33 kDa protein alone and vice versa. Thus, both cyt c-550 and the 33 kDa protein function independently in maintaining cyanobacterial oxygen-evolving activity in vivo, and both of them are required for the optimal activity. This not only provides another conclusive line of evidence for the concept that cyt c-550 functions in cyanobacterial PSII oxygen evolution but also reveals a functional difference between this extrinsic cyt in cyanobacterial PSII and the extrinsic 23 kDa protein in higher plant PSII, as the 23 kDa protein alone cannot support oxygen evolution in the absence of the 33 kDa protein.

UR - http://www.scopus.com/inward/record.url?scp=0028839495&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028839495&partnerID=8YFLogxK

M3 - Article

C2 - 7548017

AN - SCOPUS:0028839495

VL - 34

SP - 12661

EP - 12668

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 39

ER -