Cloning and Characterization of a Cdna for Algal Homologue of CP26 in the Light-Harvesting Complex II of Chlamydomonas Reinhardtii

Minagawa, Jun; Han, Kab-Cho; Dohmae, Naoshi; Takio, Koji; Inoue, Yorinao

doi:10.1007/978-94-011-3953-3_90

Cited by 8 publications

(13 citation statements)

References 10 publications

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“…The abundance of Lhca2 and Lhca5 mRNAs, encoding polypeptides of photosystem I (Elrad and Grossman 2004), showed similar decreases ( Figure 6, B and C). Expression of Lhcb5 mRNA, encoding CP26 of the minor chlorophyll a-b binding protein of photosystem II (Minagawa et al 2001;Elrad and Grossman 2004), decreased to a greater magnitude than that of Lhca2 and Lhca5 ( Figure 6D). The expression profiles of the LHC genes therefore reveal coregulation of the individual components and suggest a common regulatory mechanism in response to external stimuli.…”

Section: Resultsmentioning

confidence: 99%

Gene Expression Profiling of Flagellar Disassembly in Chlamydomonas reinhardtii

2008

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Flagella are sensory organelles that interact with the environment through signal transduction and gene expression networks. We used microarray profiling to examine gene regulation associated with flagellar length change in the green alga Chlamydomonas reinhardtii. Microarrays were probed with fluorescently labeled cDNAs synthesized from RNA extracted from cells before and during flagellar assembly or disassembly. Evaluation of the gene expression profiles identified .100 clones showing at least a twofold change in expression during flagellar length changes. Products of these genes are associated not only with flagellar structure and motility but also with other cellular responses, including signal transduction and metabolism. Expression of specific genes from each category was further characterized at higher resolution by using quantitative real-time PCR (qRT-PCR). Analysis and comparison of the gene expression profiles coupled to flagellar assembly and disassembly revealed that each process involves a new and uncharacterized whole-cell response to flagellar length changes. This analysis lays the groundwork for a more comprehensive understanding of the cellular and molecular networks regulating flagellar length changes.

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Section: Resultsmentioning

confidence: 99%

Gene Expression Profiling of Flagellar Disassembly in Chlamydomonas reinhardtii

2008

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“…Prediction of transit peptides in plant LHCII proteins by the ChloroP method was found inaccurate as well (67). The earlier attempts of CP26 amino-terminal sequencing by Edman degradation failed due to natural blocking of its amino terminus (69). It is possible that the transit peptide in the mature algal CP26 is not removed but processed by methionine excision followed by the amino-terminal acetylation as in the case of the Chlamydomonas CP29 protein (40).…”

Section: Figmentioning

confidence: 99%

Environmentally Modulated Phosphoproteome of Photosynthetic Membranes in the Green Alga Chlamydomonas reinhardtii

Turkina

Kargul

Blanco-Rivero

et al. 2006

Molecular & Cellular Proteomics

107

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Mapping of in vivoThe thylakoid membranes in chloroplasts of plants and green algae carry out oxygenic photosynthesis. Two multisubunit pigment-containing protein complexes, photosystem II (PSII) 1 and photosystem I (PSI), located in this membrane system work in series to generate an electrochemical potential gradient of protons across the membrane following vectorial electron flow from PSII to PSI via the cytochrome b 6 f complex. PSII uses light to oxidize water, whereas PSI, via a second photoact, uses reducing equivalents derived from PSII to reduce NADP ϩ to NADPH. The electrochemical potential gradient of protons is used to power conversion of ADP to ATP (1). Several thylakoid membrane proteins that make up the PSII complex and its LHCII (light-harvesting chlorophyll a/b-binding proteins of PSII) antennae undergo light-and redox-dependent phosphorylation (2, 3) as discovered more than 2 decades ago (4 -6). Phosphorylation of LHCII in plants and algae controls photosynthetic state transitions, which optimize efficient use of the absorbed light energy by both photosystems. Thus, in State 1 more energy is transferred to PSII, whereas in State 2 a proportion of the excitation energy is redistributed to PSI (7-9). The essential role of protein phosphorylation in state transitions has recently been proven in the studies using mutants of Arabidopsis thaliana plants and the green alga Chlamydomonas reinhardtii deficient in protein kinases STN7 (9) and Stt7 (8), respectively. However, despite the generally assumed similarity in thylakoid protein phosphorylation between plants and algae and a high homology between the plant STN7 and the algal Stt7 protein kinases (8), the extent of photosynthetic state transitions differs between these species. In plant thylakoids, only 15-20% of LHCII participates in the lateral migration between the photosystems, whereas up to 80% of the excitation energy absorbed by the LHCII antenna can be redistributed from PSII to PSI in green alga (8 -10).

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“…When we investigated the novel precursor proteins [11,23]), Lhcbm7 (AF479779 [11]), Lhcbm8 (AF330793. [38]), Lhcbm9 (AF479778 [11]), Lhcbm10 (AF479777 [11]), Lhcb5 (CP26 and AB050007 [32] p15, p19, and p22.2 by BLAST similarity searches, it appeared that p15 was highly similar to Lhca5, while p19 and p22.2 were most similar to Lhca2 from Arabidopsis (24) ( Table 3). To cluster the different lhca gene products, we performed phylogenetic analysis of Lhca precursor protein sequences (aligned in Fig.…”

Section: Ms (Lc-ms and Lc-ms-ms)mentioning

confidence: 99%

Proteomics of Chlamydomonas reinhardtii Light-Harvesting Proteins

et al. 2003

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With the recent development of techniques for analyzing transmembrane thylakoid proteins by two-dimensional gel electrophoresis, systematic approaches for proteomic analyses of membrane proteins became feasible. In this study, we established detailed two-dimensional protein maps of Chlamydomonas reinhardtii light-harvesting proteins (Lhca and Lhcb) by extensive tandem mass spectrometric analysis. We predicted eight distinct Lhcb proteins. Although the major Lhcb proteins were highly similar, we identified peptides which were unique for specific lhcbm gene products. Interestingly, lhcbm6 gene products were resolved as multiple spots with different masses and isoelectric points. Gene tagging experiments confirmed the presence of differentially N-terminally processed Lhcbm6 proteins. The mass spectrometric data also revealed differentially N-terminally processed forms of Lhcbm3 and phosphorylation of a threonine residue in the N terminus. The N-terminal processing of Lhcbm3 leads to the removal of the phosphorylation site, indicating a potential novel regulatory mechanism. At least nine different lhca-related gene products were predicted by comparison of the mass spectrometric data against Chlamydomonas expressed sequence tag and genomic databases, demonstrating the extensive variability of the C. reinhardtii Lhca antenna system. Out of these nine, three were identified for the first time at the protein level. This proteomic study demonstrates the complexity of the light-harvesting proteins at the protein level in C. reinhardtii and will be an important basis of future functional studies addressing this diversity.In all eukaryotic oxygenic photosynthetic organisms, lightharvesting chlorophyll a-or b-binding proteins (LHC proteins) function in the collection and transfer of light energy to the reaction centers of photosystem II (PSII) (Lhcb proteins) and photosystem I (PSI) (Lhca proteins). Additionally these proteins are also involved in light dissipation and energy quenching. Therefore, light-harvesting proteins are important components of the photosynthetic machinery that optimize photosynthetic function and minimize photooxidative damage in response to light quantity and quality. It has been known for several years that light-harvesting proteins are products of many genes. This concept is illustrated by a recent analysis of the Arabidopsis genome which revealed that the lhc gene family is composed of more than 20 genes (24). Besides the large number of lhc gene products, posttranslational modifications, such as phophorylation, contribute to even more complexity at the protein level (31, 45). Phosphorylation of the major Lhcb proteins of PSII is important in the process of state transitions. This process leads to a redistribution of excitation energy between PSII and PSI by reorganization of the antennae and thereby regulates energy flow between the photosystems. The importance of phosphorylation for state transitions is shown by the phenotype of the Chlamydomonas reinhardtii Stt7 mutant. This mutant is markedly ...

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Cloning and Characterization of a Cdna for Algal Homologue of CP26 in the Light-Harvesting Complex II of Chlamydomonas Reinhardtii

Cited by 8 publications

References 10 publications

Gene Expression Profiling of Flagellar Disassembly in Chlamydomonas reinhardtii

Gene Expression Profiling of Flagellar Disassembly in Chlamydomonas reinhardtii

Environmentally Modulated Phosphoproteome of Photosynthetic Membranes in the Green Alga Chlamydomonas reinhardtii

Proteomics of Chlamydomonas reinhardtii Light-Harvesting Proteins

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