Abstract
Heterokont algae such as diatoms and the raphidophyte Heterosigma akashiwo and peridinin-containing dinoflagellates such as Heterocapsa triquetra originally acquired their chloroplasts via secondary endosymbiosis involving a red algal endosymbiont and a eukaryote host, resulting in ‘complex’ chloroplasts surrounded by four and three membranes, respectively. The precursors of both heterokont and dinoflagellate chloroplast-targeted proteins are first inserted into the ER with removal of an N-terminal signal peptide, but how they traverse the remaining membranes is unclear. Using a nuclear-encoded thylakoid lumen protein, PsbO, from the heterokont alga Heterosigma akashiwo, the dinoflagellate Heterocapsa triquetra and the red alga Porphyra yezoensis we show that precursors without the ER signal peptide can be imported into pea chloroplasts. In the case of the H. triquetra and Porphyra PsbO, the precursors were processed to their predicted mature size and localized within the thylakoid lumen, using the Sec-dependent pathway. We report for the first time a stromal processing peptidase (SPP) activity from an alga of the red lineage. The enzyme processes the Heterosigma PsbO precursor at a single site and appears to have different substrate and reaction specificities from the plant SPP. In spite of the fact that we could not find convincing homologs of the plant chloroplast import machinery in heterokont (diatom) and red algal genomes, it is clear that these three very different lines of algae use similar mechanisms to import chloroplast precursors.
Similar content being viewed by others
Abbreviations
- CER:
-
chloroplast endoplasmic reticulum
- ER:
-
endoplasmic reticulum
- IEM and OEM:
-
inner and outer chloroplast envelope membranes
- OEC:
-
oxygen evolving complex
- PPM:
-
periplastid membrane
- PsbO:
-
33 kDa oxygen-enhancer 1 protein
- SPase:
-
signal peptidase
- SPP:
-
stromal processing peptidase
- STD:
-
stromal targeting domain
- TLCK:
-
tosyl-lysylchloromethane
- TPCK:
-
tosyl-phenylalanylchloromethane
- TPP:
-
thylakoidal processing peptidase
- TTD:
-
thylakoid targeting domain
References
M.S. Abad S.E. Clark G.K. Lamppa (1989) ArticleTitleProperties of a chloroplast enzyme that cleaves the chlorophyll a/b binding protein precursor Plant Physiol. 90 117–124
K.E. Apt N.E. Hoffman A.R. Grossman (1993) ArticleTitleThe gamma subunit of R-phycoerythrin and its possible mode of transport into the plastid of red algae J. Biol. Chem. 268 16208–16215
K.E. Apt L. Zaslavkaia J.C. Lippmeier M. Lang O. Kilian R. Wetherbee A.R. Grossman P.G. Kroth (2002) ArticleTitleIn vivo characterization of diatom multipartite plastid targeting signals J. Cell Sci. 115 4061–4069
E.V. Armbrust et al. (2004) ArticleTitleThe genome of the diatom Thalassiosira pseudonana: ecology, evolution and metabolism Science 306 79–86
D. Bhaya A. Grossman (1991) ArticleTitleTargeting proteins to diatom plastids involves transport through an endoplasmic reticulum Mol. Gen. Genet. 229 400–404
T. Cavalier-Smith (2000) ArticleTitleMembrane heredity and early chloroplast evolution Trends Plant Sci. 5 174–182
B.K. Chaal K. Ishida B.R. Green (2003) ArticleTitleA thylakoidal processing peptidase from the heterokont alga Heterosigma akashiwo Plant Mol. Biol. 52 463–472
B.K. Chaal R.M. Mould A.C. Barbrook J.C. Gray C.J. Howe (1998) ArticleTitleCharacterization of a cDNA encoding the thylakoidal processing peptidase from Arabidopsis thaliana J. Biol. Chem. 273 689–692
K. Cline (2003) Biogenesis of green plant thylakoid membranes B.R. Green W.W. Parson (Eds) Light-Harvesting Antennas in Photosynthesis Kluwer Academic Publishers Dordrecht 353–372
J.D. Dodge (1975) ArticleTitleA survey of chloroplast ultrastructure in the Dinophyceae Phycologia 14 253–263
S.P. Gibbs (1981) ArticleTitleThe chloroplast endoplasmic reticulum: structure, function, and evolutionary significance Int. Rev. Cytol. 72 49–99
B.R. Green (2004) ArticleTitleThe chloroplast genome of dinoflagellates-a reduced instruction set? Protist 155 23–31
N.R. Hofmann S.M. Theg (2003) ArticleTitlePhyscomitrella patens as a model for the study of chloroplast protein transport: conserved machineries between vascular and non-vascular plants Plant Mol. Biol. 53 621–632
K. Inoue D. Potter (2004) ArticleTitleThe chloroplastic protein translocation channel Toc75 and its paralog OEP80 represent two distinct protein families and are targeted to the chloroplastic outer envelope by different mechanisms Plant J. 39 354–365
K. Ishida T. Cavalier-Smith B.R. Green (2000) ArticleTitleEndomembrane structure and the chloroplast protein-targeting pathway in Heterosigma akashiwo (Raphidophyceae, Chromista) J. Phycol. 36 1135–1144
K. Ishida B.R. Green (2002) ArticleTitleSecond- and third-hand chloroplasts in dinoflagellates: phylogeny of oxygen-evolving enhancer 1 (PsbO) protein reveals replacement of a nuclear-encoded plastid gene by that of a haptophyte tertiary endosymbiont Proc. Natl. Acad. Sci. USA 99 9294–9299
P. Jarvis J. Soll (2002) ArticleTitleToc, Tic, and chloroplast protein import Biochim. Biophys. Acta 1590 177–189
S.W. Jeffrey G.F. Humphrey (1975) ArticleTitleNew spectrophotometric equations for determining chlorophylls a, b, c1 and c2 in higher plants, algae and natural phytoplankton Biochem. Physiol. Pflanzen. 167 191–194
S. Koussevitzky E. Ne’eman A. Sommer J.C. Steffens E. Harel (1998) ArticleTitlePurification and properties of a novel chloroplast stromal peptidase J. Biol. Chem. 273 27064–27069
P.G. Kroth (2002) ArticleTitleProtein transport into secondary plastids and the evolution of primary and secondary plastids Int. Rev. Cytol. 221 191–255
M. Lang K.E. Apt P.G. Kroth (1998) ArticleTitleProtein transport into ‘complex’ diatom plastids utilizes two different targeting signals J. Biol. Chem. 273 30973–30978
M. Matsuzaki et al. (2004) ArticleTitleGenome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D Nature 428 653–657
L. McIntosh R.A. Cattolico (1978) ArticleTitlePreservation of algal and higher plant ribosomal RNA integrity during extraction and electrophoretic quantitation Anal. Biochem. 91 600–612
D. Moreira H. Le Guyader H. Philippe (2000) ArticleTitleThe origin of red algae and the evolution of chloroplasts Nature 405 69–72 Occurrence Handle10.1038/35011054 Occurrence Handle1:STN:280:DC%2BD3c3mvFemtA%3D%3D Occurrence Handle10811219
H. Mori K. Cline (2001) ArticleTitlePost-translational protein translocation into thylakoids by the Sec and ΔpH-dependent pathways Biochim. Biophys. Acta 1541 80–90
Mould R.M. and Gray J.C. (1998). Preparation of chloroplasts for protein synthesis and protein import. In: Celis J.E. (eds) Cell Biology: A Laboratory Handbook, Academic Press, New York, pp. 81–86, 286–292
A. Nada J. Soll (2004) ArticleTitleInner envelope protein 32 is imported into chloroplasts by a novel pathway J. Cell Sci. 117 3975–3982
N. Nassoury M. Cappadocia D. Morse (2003) ArticleTitlePlastid ultrastructure defines the protein import pathway in dinoflagellates J. Cell Sci. 116 2867–2874
H. Nielsen J. Engelbrecht S. Brunak G. Heijne Particlevon (1997) ArticleTitleIdentification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites Protein Eng. 10 1–6 Occurrence Handle10.1093/protein/10.1.1
I. Nikaido E. Asamizu M. Nakajima Y. Nakamura N. Saga S. Tabata (2000) ArticleTitleGeneration of 10,154 expressed sequence tags from a leafy gametophyte of a marine red alga, Porphyra yezoensis DNA Res. 7 223–227
S. Reinbothe F. Quigley A. Springer A. Schemenewitz C. Reinbothe (2004) ArticleTitleThe outer plastid envelope protein Oep16: role as precursor translocase in import of protochlorophyllide oxidoreductase A Proc. Natl. Acad. Sci. USA 101 2203–2208
S. Reumann K. Keegstra (1999) ArticleTitleThe endosymbiotic origin of the protein import machinery of chloroplastic envelope membranes Trends Plant Sci. 4 302–307
S. Richter G.K. Lamppa (1998) ArticleTitleA chloroplast processing enzyme functions as the general stromal processing peptidase Proc. Natl. Acad. Sci. USA 95 7463–7468
S. Richter G.K. Lamppa (2003) ArticleTitleStructural properties of the chloroplast stromal processing peptidase required for its function in transit peptide removal J. Biol. Chem. 278 39497–39502
A. Rüfenacht A. Boschetti (2000) ArticleTitleChloroplasts of the green alga Chlamydomonas reinhardtii possess at least four distinct stromal processing proteases Photosynth. Res. 63 249–258
J. Sambrook E.F. Fritsch T. Maniatis (1989) Molecular Cloning: A Laboratory Manual EditionNumber2 Cold Spring Harbor Laboratory Press New York
C.D. Scaramuzzi R.G. Hiller H.W. Stokes (1992) ArticleTitleIdentification of a chloroplast-encoded secA gene homologue in a chromophytic alga: possible role in chloroplast protein translocation Curr. Genet. 22 421–427
M. Schubert U.A. Petersson B.J. Haas C. Funk W.P. Schröder T. Kieselbach (2002) ArticleTitleProteome map of the chloroplast lumen of Arabidopsis thaliana J. Biol. Chem. 277 8354–8365
J. Soll E. Schleiff (2004) ArticleTitleProtein import into chloroplasts Nat. Rev. Mol Cell Biol. 5 198–208
Q. Su A. Boschetti (1993) ArticleTitlePartial purification and properties of enzymes involved in the processing of a chloroplast import protein from Chlamydomonas reinhardtii Eur. J. Biochem. 217 1039–1047
K. Valentin (1997) ArticleTitlePhylogeny and expression of the secA gene from a chromophytic alga - implications for the evolution of plastids and sec-dependent protein translocation Curr. Genet. 32 300–307
Zhang X-P. E. Glaser (2002) ArticleTitleInteraction of plant mitochondrial and chloroplast signal peptides with the Hsp70 molecular chaperone Trends Plant Sci. 7 14–21
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chaal, B.K., Green, B.R. Protein import pathways in ‘complex’ chloroplasts derived from secondary endosymbiosis involving a red algal ancestor. Plant Mol Biol 57, 333–342 (2005). https://doi.org/10.1007/s11103-004-7848-y
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11103-004-7848-y