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Understanding Photosynthetic Electron Transport Using Chlamydomonas: The Path from Classical Genetics to High Throughput Genomics

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Book cover Functional Genomics and Evolution of Photosynthetic Systems

Summary

The Volvocales, an order of the green algal class Chlorophyceae, and the Streptophyte algae, the lineage that evolved into land plants, shared a common ancestor about one billion years ago. Chlamydomonas reinhardtii (Chlamydomonas throughout) a unicellular member of the Volvocales, has traditionally been considered a strong model organism that has been probed with sophisticated tools and methodologies to elucidate numerous biological processes. Perhaps the most in-depth analyses of Chlamydomonas have focused on defining proteins and complexes involved in the function and biogenesis of chloroplasts as well as the structure, assembly, and function of eukaryotic flagella (cilia); the latter are inherited from the common ancestor of animals and plants, but were lost during the evolution of land plants. This review emphasizes how Chlamydomonas has been used to elucidate a number of different activities associated with photosynthetic function. Many of these analyses were performed using classical genetic, biochemical and physiological approaches. However, recently, the DOE – Joint Genome Institute has sequenced the nuclear genome of Chlamydomonas (∼120 Mb) and has helped the community of researchers perform comparative genomic analyses. Comparisons of deduced Chlamydomonas proteins has identified a set of proteins specifically present in the green lineage and photosynthetic organisms, but not present in nonphotosynthetic organisms; this protein assemblage has been designated the GreenCut. Many proteins in the GreenCut are likely resident in the chloroplast and potentially associated with photosynthetic processes. Toward the end of this text we discuss the ways in which genomics has added a new dimension to our analyses of photosynthetic processes.

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Abbreviations

A:

antheroxanthin

ALAD:

δ-aminolevulinic acid dehydratase

BAC:

bacterial artificial chromosome

CES:

control of epistasis of synthesis

CGL:

refers to proteins of the GreenCut conserved in the green lineage organisms

CGLD:

refers to proteins of the GreenCut conserved in the green lineage organisms and the diatoms

Chl:

chlorophyll

Chl+ :

chlorophyll cation

CHLH1:

H subunit of Mg-chelatase

CHLI1:

I subunit of the Mg-chelatase

CTH1:

catalyzes the cyclase reaction in chlorophyll biosynthesis

CP43:

chlorophyll binding antenna protein tightly bound to photosystem II

CPXI:

coproporphyrinogen oxidase

CRD:

an iron requiring cyclase involve in chlorophyll synthesis

D1:

the rapidly turning over reaction center protein of photosystem II

D2:

reaction center protein of photosystem II that associates with D1

DEG:

protease involved in chloroplast biogenesis

Fd:

ferredoxin

Fe:

iron

FNR:

ferredoxin NADP oxido-reductase

FQR:

ferredoxin-quinone reductase

FLU:

protein that regulates chlorophyll biosynthesis

EST:

expressed sequence tag

FTSH:

protease involved in the turnover of proteins of the photosynthetic apparatus

GSA:

glutamate 1-semialdehyde (GSA) aminotransferase

GLK:

golden-like kinase transcriptional regulator

HEMA:

glutamyl tRNA reductase

IsiA:

antenna chlorophyll binding proteins synthesized during iron deprivation (similar to CP43)

IsiB:

flavodoxin synthesized during iron deprivation

JGI:

Joint Genome Institute

LHC:

light harvesting complex

LHCII:

light harvesting complex II

LHCA:

light harvesting complex of photosystem I

LHCB:

light harvesting proteins associated with photosystem II

LHCSR:

protein in the light harvesting complex family that may be involved in photoprotection in Chlamydomonas

MCA1:

protein required for stable accumulation of petA RNA

Mg-ProtoIX:

Mg-protoporphyrin IX

Mg-ProtoIXMe:

Mg-protoporphyrin IX-monomethylester

NAB1:

forms a complex with mRNA encoding light harvesting proteins

NDH1:

NADH:ubiquinone oxidoreductase

OEE:

oxygen evolving complex

PGR1 and PGR5:

protein thought to be part of the ferredoxin-quinone reductase complex

NPQ:

non-photochemical quenching

PetA:

cytochrome f

PSI:

photosystem I

PSII:

photosystem II

PSAH or PSIH:

specific polypeptide associated with photosystem II

PSA:

proteins associated with photosystem I (an additional letter indicates the subunit of the complex)

PSB:

protein associated with photosystem II (an additional letter indicates the subunit of the complex)

PSBS:

protein in the light harvesting family involved in qE-based quenching

PQ:

plastoquinone

qE:

quenching through the formation of an electrochemical gradient

qI:

quenching through inhibition of photosystem II

qT:

quenching through the formation of a state transition

rbcL :

gene encoding the large subunit of ribulose-1,5-bisphosphate carboxylase

RNAi:

RNA interference

RNA-seq:

new generation of RNA sequencing

ROS:

reactive oxygen species

RuBP:

carboxylase ribulose-1,5-bisphosphate carboxylase

STN7:

Arabidopsis serine threonine protein kinase associated with state transition

STN8:

Arabidopsis protein with homology to STN7

STT7:

Chlamydomonas serine threonine protein kinase associated with state transition

TCA1:

protein involved in the translation of petA mRNA

TIC:

proteins on the inner membrane of the chloroplast envelop involved in transporting proteins into the chloroplast

TOC:

proteins on outer inner membrane of the chloroplast envelop involved in transporting proteins into the chloroplast

V:

violaxanthin

Z:

zeaxanthin

Z+ :

zeaxanthin cation

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Acknowledgments

Much of the genome analysis discussed in this manuscript could not have been performed without the help of Dan Rokhsar and Simon Prochnik, who are responsible for the Chlamydomonas genome project at DOE’s Joint Genome Institute. Aspects of the work presented were also supported by NSF grants MCB0235878, MCB0951094 and Department of energy grant DE-FG0207ER64427 awarded to ARG and by the Marie Curie OIF-6 APOSD grant (EU). SM acknowledges support from the Department of Energy (DE-FC03-02ER63421 and DE-FG02-07ER15229). SJK was supported in part by a Ruth L. Kirschstein National Research Service Award GM07185.

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Grossman, A.R., González-Ballester, D., Bailey, S., Karpowicz, S.J., Merchant, S.S. (2012). Understanding Photosynthetic Electron Transport Using Chlamydomonas: The Path from Classical Genetics to High Throughput Genomics. In: Burnap, R., Vermaas, W. (eds) Functional Genomics and Evolution of Photosynthetic Systems. Advances in Photosynthesis and Respiration, vol 33. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-1533-2_6

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