title: Protein import pathways in 'complex' chloroplasts derived from secondary endosymbiosis involving a red algal ancestor.
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.
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, Canada.
authors.forename: Balbir K
authors.initials: BK
authors.lastname: Chaal
authors.forename: Beverley R
authors.initials: BR
authors.lastname: Green
mesh_terms: D020418:Algal Proteins / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002736:Chloroplasts / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004141:Dinoflagellida / Q000378:metabolism; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000201:enzymology; D008565:Membrane Proteins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D021381:Protein Transport / Q000187:drug effects; D015800:Protozoan Proteins / Q000378:metabolism; D000461:Rhodophyta / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000378:metabolism; D019810:Sodium Azide / Q000494:pharmacology; D013559:Symbiosis; D013820:Thermolysin / Q000494:pharmacology; D020524:Thylakoids / Q000187:drug effects / Q000378:metabolism
chemical_list: D020418:Algal Proteins; D008565:Membrane Proteins; D010940:Plant Proteins; D015800:Protozoan Proteins; D019810:Sodium Azide; D012697:Serine Endopeptidases; C026515:type I signal peptidase; D008666:Metalloendopeptidases; C071986:stromal processing peptidase; D013820:Thermolysin