Protein targeting into complex diatom plastids: functional characterization of a specific targeting motif

Gruber, Ansgar; Vugrinec, Sascha; Hempel, Franziska; Gould, Sven B.; Maier, Uwe-G.; Kroth, Peter G.

Protein targeting into complex diatom plastids: functional characterization of a specific targeting motif

Dateien

Gruber_suppl_mat.pdfGröße: 224.38 KBDownloads: 479

Gruber_Protein_Targeting.pdfGröße: 609.7 KBDownloads: 538

Datum

2007

Autor:innen

Gruber, Ansgar

Vugrinec, Sascha

Hempel, Franziska

Gould, Sven B.

Maier, Uwe-G.

Kroth, Peter G.

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Plant Molecular Biology. 2007, 64(5), pp. 519-530. ISSN 0167-4412. eISSN 1573-5028. Available under: doi: 10.1007/s11103-007-9171-x

Zusammenfassung

Plastids of diatoms and related algae evolved by secondary endocytobiosis, the uptake of a eukaryotic alga into a eukaryotic host cell and its subsequent reduction into an organelle. As a result diatom plastids are surrounded by four membranes. Protein targeting of nucleus encoded plastid proteins across these membranes depends on N-terminal bipartite presequences consisting of a signal and a transit peptide-like domain. Diatoms and cryptophytes share a conserved amino acid motif of unknown function at the cleavage site of the signal peptides (ASAFAP), which is particularly important for successful plastid targeting. Screening genomic databases we found that in rare cases the very conserved phenylalanine within the motif may be replaced by tryptophan, tyrosine or leucine. To test such unusual presequences for functionality and to better understand the role of the motif and putative receptor proteins involved in targeting, we constructed presequence: GFP fusion proteins with or without modifications of the "ASAFAP"-motif and expressed them in the diatom Phaeodactylum tricornutum. In this comprehensive mutational analysis we found that only the aromatic amino acids phenylalanine, tryptophan, tyrosine and the bulky amino acid leucine at the +1 position of the predicted signal peptidase cleavage site allow plastid import, as expected from the sequence comparison of native plastid targeting presequences of P. tricornutum and the cryptophyte Guillardia theta. Deletions within the signal peptide domains also impaired plastid import, showing that the presence of F at the N-terminus of the transit peptide together with a cleavable signal peptide is crucial for plastid import.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

Chloroplast, Diatom, Evolution, Import, Presequence

Zitieren

ISO 690

GRUBER, Ansgar, Sascha VUGRINEC, Franziska HEMPEL, Sven B. GOULD, Uwe-G. MAIER, Peter G. KROTH, 2007. Protein targeting into complex diatom plastids: functional characterization of a specific targeting motif. In: Plant Molecular Biology. 2007, 64(5), pp. 519-530. ISSN 0167-4412. eISSN 1573-5028. Available under: doi: 10.1007/s11103-007-9171-x

BibTex

@article{Gruber2007Prote-7070,
  year={2007},
  doi={10.1007/s11103-007-9171-x},
  title={Protein targeting into complex diatom plastids: functional characterization of a specific targeting motif},
  number={5},
  volume={64},
  issn={0167-4412},
  journal={Plant Molecular Biology},
  pages={519--530},
  author={Gruber, Ansgar and Vugrinec, Sascha and Hempel, Franziska and Gould, Sven B. and Maier, Uwe-G. and Kroth, Peter G.}
}

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Universitätsbibliographie

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