Protein targeting of an unusual, evolutionarily conserved adenylate kinase to a eukaryotic flagellum.
The eukaryotic flagellum is a large structure into which specific constituent proteins must be targeted, transported and assembled after their synthesis in the cytoplasm. Using Trypanosoma brucei and a proteomic approach, we have identified and characterized a novel set of adenylate kinase proteins that are localized to the flagellum. These proteins represent unique isoforms that are targeted to the flagellum by an N-terminal extension to the protein and are incorporated into an extraaxonemal structure (the paraflagellar rod). We show that the N-terminal extension is both necessary for isoform location in the flagellum and sufficient for targeting of a green fluorescent protein reporter protein to the flagellum. Moreover, these N-terminal extension sequences are conserved in evolution and we find that they allow the identification of novel adenylate kinases in the genomes of humans and worms. Given the existence of specific isoforms of certain central metabolic enzymes, and targeting sequences for these isoforms, we suggest that these isoforms form part of a complex, "solid-phase" metabolic capability that is built into the eukaryotic flagellum.
Pullen
Timothy J
TJ
Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.
Ginger
Michael L
ML
Gaskell
Simon J
SJ
Gull
Keith
K
D000263:Adenylate Kinase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D019143:Evolution, Molecular; D005407:Flagella / Q000201:enzymology* / Q000276:immunology / Q000648:ultrastructure; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D040901:Proteomics; D034741:RNA, Small Interfering / Q000494:pharmacology; D016415:Sequence Alignment; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D007527:Isoenzymes; D034741:RNA, Small Interfering; D000263:Adenylate Kinase