Abstract
Main conclusion
l-Arginine supports growth and resulted in increased PII signaling protein levels and lipid droplet accumulation in the colorless green alga Polytomella parva.
Abstract
Polytomella parva, a model system for nonphotosynthetic green algae, utilizes ammonium and several carbon sources, including ethanol and acetate. We previously reported that P. parva accumulates high amounts of arginine with the key enzyme of the ornithine/arginine biosynthesis pathway, N-acetyl-L-glutamate kinase, exhibiting high activity. Here we demonstrate that l-arginine can be used by this alga as a nitrogen source. Externally supplied arginine directly influenced the levels of PII signaling protein and formation of triacylglycerol (TAG)-filled lipid bodies (LBs). Our results suggest that the nitrogen source, but not nitrogen starvation, may be critical for the accumulation of LBs in a PII-independent manner in P. parva.
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Abbreviations
- LBs:
-
Lipid bodies
- NAGK:
-
N-Acetyl-l-glutamate kinase
- TAG:
-
Triacylglycerol
References
Atteia A, van Lis R, Ramírez J, González-Halphen D (2000) Polytomella spp. growth on ethanol. Extracellular pH affects the accumulation of mitochondrial cytochrome c550. Eur J Biochem 267(10):2850–2858
Chellamuthu VR, Ermilova E, Lapina T, Lüddecke J, Minaeva E, Herrmann C, Hartmann MD, Forchhammer K (2014) A widespread glutamine-sensing mechanism in the plant kingdom. Cell 159(5):1188–1199. https://doi.org/10.1016/j.cell.2014.10.015
Ermilova E, Lapina T, Zalutskaya Z, Minaeva E, Fokina O, Forchhammer K (2013) PII signal transduction protein in Chlamydomonas reinhardtii: localization and expression pattern. Protist 164:49–59
Feria Bourrellier AB, Valot B, Ambard-Bretteville F, Vidal J, Hodges M (2010) Chloroplast acetyl-CoA carboxylase activity is 2-oxoglutarate-regulated by interaction of PII with the biotin carboxyl carrier subunit. Proc Natl Acad Sci USA 107:502–507. https://doi.org/10.1073/pnas.0910097107
Ferrario-Méry S, Bouvet M, Leleu O, Savino G, Hodges M, Meyer C (2005) Physiological characterization of Arabidopsis mutants affected in the expression of the putative regulatory protein PII. Planta 223:28–39. https://doi.org/10.1007/s00425-005-0063-5
Gerhardt ECM, Rodrigues TE, Müller-Santos M, Pedrosa FO, Souza EM, Forchhammer K, Huergo LF (2014) The Bacterial signal transduction protein GlnB regulates the committed step in fatty acid biosynthesis by acting as a dissociable regulatory subunit of acetyl-CoA carboxylase. Mol Microbiol 95(6):1025–1035. https://doi.org/10.1111/mmi.12912
Giordano M, Raven JA (2014) Nitrogen and sulfur assimilation in plants and algae. Aquat Bot 118:45–61. https://doi.org/10.1016/j.aquabot.2014.06.012
Greenspan P, MayerEP FowlerSD (1985) Nile red: a selective fluorescent stain for intracellular lipid droplets. JCB 100:965–973
Grossman A, Takahashi H (2001) Macronutrient utilization by photosynthetic eukaryotes and the fabric of interactions. Annu Rev Plant Physiol 52:163–210
Han R, Khalid M, Juan J, Huang D (2018) Exogenous glycine inhibits root elongation and reduces nitrate-N uptake in pak choi (Brassica campestris ssp. Chinensis L.). PLoS ONE 13(9):e0204488. https://doi.org/10.1371/journal.pone.0204488
Hauf W, Schmid K, Gerhardt EC, Huergo LF, Forchhammer K (2016) Interaction of the nitrogen regulatory protein GlnB (PII) with biotin carboxyl carrier protein (BCCP) controls acetyl-CoA levels in the cyanobacterium Synechocystis sp. PCC 6803. Front Microbiol 7:1700. https://doi.org/10.3389/fmicb.2016.01700
Heinrich A, Maheswaran M, Ruppert U, Forchhammer K (2004) The Synechococcus elongatus P signal transduction protein controls arginine synthesis by complex formation with N-acetyl-L-glutamate kinase. Mol Microbiol 52(5):1303–1314. https://doi.org/10.1111/j.1365-2958.2004.04058.x
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lapina T, Selim KA, Forchhammer K, Ermilova E (2018) The PII signaling protein from red algae represents an evolutionary link between cyanobacterial and Chloroplastida PII proteins. Sci Rep 8(1):790. https://doi.org/10.1038/s41598-017-19046-7
Minaeva E, Ermilova E (2015) Sequencing and expression analysis of the gene encoding PII signal protein in Chlorella variabilis NC64A. J Plant Biochem Physiol 3:142. https://doi.org/10.4172/2329-9029.1000142
Minaeva E, Forchhammer K, Ermilova E (2015) Glutamine assimilation and feedback regulation of L-acetyl-N-glutamate kinase activity in Chlorella variabilis NC64A results in changes in arginine pools. Protist 166:493–505. https://doi.org/10.1016/j.protis.2015.08.001
Näsholm T, Kielland K, Ganeteg U (2009) Uptake of organic nitrogen by plants. New Phytol 182(1):31–48. https://doi.org/10.1111/j.1469-8137.2008.02751.x
Popov N, Schmitt M, Schulzeck S, Matthies H (1975) Reliable micromethod for determination of the protein content in tissue homogenates. Acta Biol Med Ger 34:1441–1446
Rodrigues TE, Gerhardt EC, Oliveira MA, Chubatsu LS, Pedrosa FO, Souza EM, Souza GA, Müller-Santos M, Huergo LF (2014) Search for novel targets of the PII signal transduction protein in bacteria identifies the BCCP component of acetyl-CoA carboxylase as a PII binding partner. Mol Microbiol 91:751–761. https://doi.org/10.1111/mmi.12493
Sakaguchi S (1950) A new method for the colorimetric determination of arginine. J Biochem 37:231–236
Sanz-Luque E, Chamizo-Ampudia A, Llamas A, Galvan A, Fernandez E (2015) Understanding nitrate assimilation and its regulation in microalgae. Front Plant Sci 6:1–17. https://doi.org/10.3389/fpls.2015.00899
Selim K, Lapina T, Forchhammer K, Ermilova E (2019) Interaction of N-acetyl-L-glutamate kinase with the PII signal transducer in the nonphotosynthetic alga Polytomella parva: co-evolution towards a hetero-oligomeric enzyme. FEBS J. https://doi.org/10.1111/febs.14989
Smith DR, Lee RW (2014) A plastid without a genome: evidence from the non photosynthetic green algal genus Polytomella. Plant Physiol 164(4):1812–1819. https://doi.org/10.1104/pp.113.233718
Sugiyama K, Hayakawa T, Kudo T, Ito T, Yamaya T (2004) Interaction of N-acetyl glutamate kinase with a PII-like protein in rice. Plant Cell Physiol 45:1768–1778. https://doi.org/10.1093/pcp/pch199
Wise DL (1955) Carbon sources for Polytomella caeca. J Protozool 2:156–158
Zalutskaya Zh, Kharatyan N, Forchhammer K, Ermilova E (2015) Reduction of PII signaling protein enhances lipid body production in Chlamydomonas reinhardtii. Plant Sci 240:1–9. https://doi.org/10.1016/j.plantsci.2015.08.019
Acknowledgements
This work was supported by Saint-Petersburg State University (Grant no. 1.65.38.2017) and by the Russian Science Foundation (Grant no.16-14-10004-П) to E.E., and DFG to K.F. (Fo195/9-2, Fo195/13-1).
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Lapina, T.V., Kochemasova, L.Y., Forchhammer, K. et al. Effects of arginine on Polytomella parva growth, PII protein levels and lipid body formation. Planta 250, 1379–1385 (2019). https://doi.org/10.1007/s00425-019-03249-5
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DOI: https://doi.org/10.1007/s00425-019-03249-5