Plastid proteome prediction for diatoms and other algae with secondary plastids of the red lineage.

Ansgar Gruber,Peter G Kroth,Gabrielle Rocap,E Virginia Armbrust,Thomas Mock

doi:10.1111/tpj.12734

Ansgar Gruber, Peter G Kroth + Show 3 more

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https://doi.org/10.1111/tpj.12734

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Abstract

The plastids of ecologically and economically important algae from phyla such as stramenopiles, dinoflagellates and cryptophytes were acquired via a secondary endosymbiosis and are surrounded by three or four membranes. Nuclear-encoded plastid-localized proteins contain N-terminal bipartite targeting peptides with the conserved amino acid sequence motif ‘ASAFAP’. Here we identify the plastid proteomes of two diatoms, Thalassiosira pseudonana and Phaeodactylum tricornutum, using a customized prediction tool (ASAFind) that identifies nuclear-encoded plastid proteins in algae with secondary plastids of the red lineage based on the output of SignalP and the identification of conserved ‘ASAFAP’ motifs and transit peptides. We tested ASAFind against a large reference dataset of diatom proteins with experimentally confirmed subcellular localization and found that the tool accurately identified plastid-localized proteins with both high sensitivity and high specificity. To identify nucleus-encoded plastid proteins of T. pseudonana and P. tricornutum we generated optimized sets of gene models for both whole genomes, to increase the percentage of full-length proteins compared with previous assembly model sets. ASAFind applied to these optimized sets revealed that about 8% of the proteins encoded in their nuclear genomes were predicted to be plastid localized and therefore represent the putative plastid proteomes of these algae.

Highlights

Plastids arose through endosymbiotic processes – a primary endosymbiosis of a cyanobacterium gave rise to red and green algae and the subsequent evolution of plants, and multiple secondary endosymbioses of either a red or a green alga gave rise to a broad diversity of eukaryotic microbes
The plastid protein prediction was initiated with a set of putative plastid-targeted proteins from the diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum, compiled based on the lists of nucleus-encoded and plastid-targeted proteins published by Armbrust et al (2004), Gruber et al (2007), and Kroth et al (2008) (P. tricornutum and T. pseudonana)
To avoid potential overfitting of the data, we reduced the level of homology within the protein set using an ‘all against all’ BLAST search of the candidate sequences from T. pseudonana and P. tricornutum

Summary

SUMMARY

The plastids of ecologically and economically important algae from phyla such as stramenopiles, dinoflagellates and cryptophytes were acquired via a secondary endosymbiosis and are surrounded by three or four membranes. Nuclear-encoded plastid-localized proteins contain N-terminal bipartite targeting peptides with the conserved amino acid sequence motif ‘ASAFAP’. We identify the plastid proteomes of two diatoms, Thalassiosira pseudonana and Phaeodactylum tricornutum, using a customized prediction tool (ASAFind) that identifies nuclear-encoded plastid proteins in algae with secondary plastids of the red lineage based on the output of SignalP and the identification of conserved ‘ASAFAP’ motifs and transit peptides. To identify nucleus-encoded plastid proteins of T. pseudonana and P. tricornutum we generated optimized sets of gene models for both whole genomes, to increase the percentage of full-length proteins compared with previous assembly model sets. ASAFind applied to these optimized sets revealed that about 8% of the proteins encoded in their nuclear genomes were predicted to be plastid localized and represent the putative plastid proteomes of these algae

INTRODUCTION

RESULTS AND DISCUSSION

Results of prediction no

CONCLUSION

EXPERIMENTAL PROCEDURES