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FEBS Letters 582 (2008) 1413–1418
Parkin-co-regulated gene (PACRG) product interacts with tubulin and microtubules
Takashi Ikeda*,1 Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Received 7 December 2007; revised 18 February 2008; accepted 25 February 2008
Available online 1 April 2008
Edited by Judit Ova´di
tility in mouse [5] and in the stable assembly of axonemal outer Abstract Parkin-co-regulated gene (PACRG) is a gene that shares a bidirectional promoter with ParkinsonÕs disease-related doublet microtubules in Trypanosoma [6]. Our study of Chla- Parkin/Park2 gene. Recently, the PACRG gene product was mydomonas PACRG showed that it is densely localized along implicated in the function of flagella. However, its exact function the flagellar outer doublet microtubules, possibly buried in the remains unknown. Here, I assessed the interaction between microtubule wall [7]. Despite these findings, the biochemical PACRG and tubulin. Co-sedimentation experiments revealed nature and physiological function of PACRG remain to be that PACRG directly binds to microtubules and a/b-tubulin het- studied. Here, I report that PACRG associates with both erodimers with high affinity. Microscopic studies showed that microtubules and tubulin dimers in vitro. PACRG bundles microtubules and forms branched aggregates with unpolymerized tubulin dimers. The amino acid sequence of the microtubule-binding region of PACRG is highly conserved among various organisms, suggesting that tubulin binding is a 2. Materials and methods basic property of PACRG. Structured summary: 2.1. Alignment By BLAST search using the amino acid sequence of C. reinhardtii MINT-6439731, MINT-6439753, MINT-6439768: PACRG (GenBank Accession No. BAG12189) [7] as the query, PACRG (genbank_protein_gi:169219363), tubulin alpha (uni- homologous sequences were obtained from T. brucei (XP_843825, protkb:P02550), tubulin beta (uniprotkb:Q2XVP4) bind XP_827292), Paramecium tetraurelia (CAK71037, XP_001443020), (MI:0407) by cosedimentation (MI:0027) Caenorhabditis elegans (NP_495496), Tetrahymena thermophila MINT-6439796: (XP_001023392, XP_001022215, XP_001022189), Strongylocentrotus purpuratus (XP_791188, XP_784874), Drosophila melanogaster PACRG (genbank_protein_gi:169219363), tubulin alpha (uni- (NP_609957, XP_843825), Apis mellifera (XP_625146), Danio rerio protkb:P02550), tubulin beta (uniprotkb:Q2XVP4) bind (NP_001007399), Mus musculus (NP_081308), and Homo sapiens (MI:0407) by pull-down (MI:0096) (AAH30642). Alignment was created using CLUSTALW ver. 1.83 [8].