Kinase-associated Endopeptidase 1 (Kae1) Participates in an Atypical Ribosome-associated Complex in the Apicoplast of Plasmodium falciparum
| Autor: | Anna Oksman, Jeremy P. Mallari, Barbara A. Vaupel, Daniel E. Goldberg |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
TRNA modification
Protein family Immunoprecipitation Molecular Sequence Data Plasmodium falciparum Protozoan Proteins Apicoplasts Biochemistry Ribosome Mass Spectrometry Conserved sequence Cytosol Nucleic Acids Animals Cluster Analysis Humans Parasites Amino Acid Sequence Protein Interaction Maps Molecular Biology Alleles Conserved Sequence Phylogeny Life Cycle Stages Apicoplast biology Molecular Sequence Annotation Cell Biology DNA Protozoan biology.organism_classification Recombinant Proteins Cell Compartmentation Cell biology Transport protein Protein Transport RNA Ribosomal Ribosomes Sequence Alignment Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 289:30025-30039 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m114.586735 |
| Popis: | The universally conserved kinase-associated endopeptidase 1 (Kae1) protein family has established roles in N(6)-threonylcarbamoyl adenosine tRNA modification, transcriptional regulation, and telomere homeostasis. These functions are performed in complex with a conserved core of protein binding partners. Herein we describe the localization, essentiality, and protein-protein interactions for Kae1 in the human malaria parasite Plasmodium falciparum. We found that the parasite expresses one Kae1 protein in the cytoplasm and a second protein in the apicoplast, a chloroplast remnant organelle involved in fatty acid, heme, and isoprenoid biosynthesis. To analyze the protein interaction networks for both Kae1 pathways, we developed a new proteomic cross-validation approach. This strategy compares immunoprecipitation-MS data sets across different cellular compartments to enrich for biologically relevant protein interactions. Our results show that cytoplasmic Kae1 forms a complex with Bud32 and Cgi121 as in other organisms, whereas apicoplast Kae1 makes novel interactions with multiple proteins in the apicoplast. Quantitative RT-PCR and immunoprecipitation studies indicate that apicoplast Kae1 and its partners interact specifically with the apicoplast ribosomes and with proteins involved in ribosome function. Together, these data indicate an expanded, apicoplast-specific role for Kae1 in the parasite. |
| Databáze: | OpenAIRE |
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