Kinetic and phylogenetic analysis of plant polyamine uptake transporters
The rice gene POLYAMINE UPTAKE TRANSPORTER1 (PUT1) was originally identified based on its homology to the polyamine uptake transporters LmPOT1 and TcPAT12 in Leishmania major and Trypanosoma cruzi, respectively. Here we show that five additional transporters from rice and Arabidopsis that cluster in the same clade as PUT1 all function as high affinity spermidine uptake transporters. Yeast expression assays of these genes confirmed that uptake of spermidine was minimally affected by 166 fold or greater concentrations of amino acids. Characterized polyamine transporters from both Arabidopsis thaliana and Oryza sativa along with the two polyamine transporters from L. major and T. cruzi were aligned and used to generate a hidden Markov model. This model was used to identify significant matches to proteins in other angiosperms, bryophytes, chlorophyta, discicristates, excavates, stramenopiles and amoebozoa. No significant matches were identified in fungal or metazoan genomes. Phylogenic analysis showed that some sequences from the haptophyte, Emiliania huxleyi, as well as sequences from oomycetes and diatoms clustered closer to sequences from plant genomes than from a homologous sequence in the red algal genome Galdieria sulphuraria, consistent with the hypothesis that these polyamine transporters were acquired by horizontal transfer from green algae. Leishmania and Trypansosoma formed a separate cluster with genes from other Discicristates and two Entamoeba species. We surmise that the genes in Entamoeba species were acquired by phagotrophy of Discicristates. In summary, phylogenetic and functional analysis has identified two clades of genes that are predictive of polyamine transport activity.
KeywordsArabidopsis Hidden Markov model Michaelis–Menten Oryza PUT1 Spermidine
Hidden Markov model
Multiple Em for Motif Elicitation
This work was supported by grants from the Ohio Plant Biotechnology Consortium, and the Office of Sponsored Programs at BGSU.
Conflict of interest
The authors declare that they have no conflict of interest.
- Alet AI, Sanchez DH, Cuevas JC, Del Valle S, Altabella T, Tiburcio AF, Marco F, Ferrando A, Espasandin FD, Gonzalez ME, Ruiz OA, Carrasco P (2011) Putrescine accumulation in Arabidopsis thaliana transgenic lines enhances tolerance to dehydration and freezing stress. Plant Signal Behav 6:278–286PubMedCrossRefGoogle Scholar
- Antognoni F, Pistocchi R, Bagni N (1993) Uptake competition between polyamines and analogues in carrot protoplasts. Plant Physiol Biochem 31:693–698Google Scholar
- Aurrecoechea C, Brestelli J, Brunk BP, Fischer S, Gajria B, Gao X, Gingle A, Grant G, Harb OS, Heiges M, Innamorato F, Iodice J, Kissinger JC, Kraemer ET, Li W, Miller JA, Nayak V, Pennington C, Pinney DF, Roos DS, Ross C, Srinivasamoorthy G, Stoeckert CJ Jr, Thibodeau R, Treatman C, Wang H (2010) EuPathDB: a portal to eukaryotic pathogen databases. Nucleic Acids Res 38(Database issue):D415–D419Google Scholar
- Baldauf SL (2008) An overview of the phylogeny and diversity of eukaryotes. J Syst Evol 46:263–275Google Scholar
- Cohen SS (1998) A guide to the polyamines. Oxford University Press New York, 595 ppGoogle Scholar
- Cuevas J, Lopez-Cobollo R, Alcazar R, Zarza X, Koncz C, Altabella T, Salinas J, Tiburcio A, Ferrando A (2008) Putrescine is involved in Arabidopsis freezing tolerance and cold acclimation by regulating abscisic acid levels in response to low temperature. Plant Physiol 148:1094–1105PubMedCrossRefGoogle Scholar
- Moschou P, Paschalidis K, Delis I, Andriopoulou A, Lagiotis G, Yakoumakis D, Roubelakis-Angelakis K (2008) Spermidine exodus and oxidation in the apoplast induced by abiotic stress is responsible for H2O2 signatures that direct tolerance responses in tobacco. Plant Cell 20:1708–1724PubMedCrossRefGoogle Scholar
- Neily MH, Matsukura C, Maucourt M, Bernillon S, Deborde C, Moing A, Yin YG, Saito T, Mori K, Asamizu E, Rolin D, Moriguchi T, Ezura H (2011) Enhanced polyamine accumulation alters carotenoid metabolism at the transcriptional level in tomato fruit over-expressing spermidine synthase. J Plant Physiol 168:242–252PubMedCrossRefGoogle Scholar