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Polyamines are well known for their wide range of roles in the cells, including stabilization of nucleic acids, regulation of gene expression at different levels, stress response and cell cycle. The activity of these well conserved small polycations are also regulated by different mechanisms to assure their intracellular concentration and activity. Polyamine transporters have a major role in the regulation of the influx and efflux of the polyamines. Previously, two spermine family transporters SPBC409.08 and caf5 were characterized which were associated with cell size defects. The aim of this study is to better understand the role of caf5 plasma membrane spermine transporter and its functional interaction with spermidine transporters. Double mutants of caf5 and spermidine family transporters SPBC36.01c, SPBC36.03c and SPBC569.05c were formed in S. pombe. The double mutants were then put through growth rate analysis, meiotic progression and sporulation analysis, cell cycle analysis, and vulnerability to environmental stress. The results showed that the double mutants differentiate from each other in their combined mutation with caf5 in terms of cell size and DNA damage response, indicating redundant seperate pathways of spermine and spermidine transport in S. pombe.

Anahtar Kelimeler:

S. pombe, Cell Size, Cell Cycle, Stress Response, Polyamines

FUNCTIONAL INTERACTION BETWEEN caf5 AND SPERMIDINE FAMILY TRANSPORTERS IN S. POMBE

ABSTRACT Polyamines are well known for their wide range of roles in the cells, including stabilization of nucleic acids, regulation of gene expression at different levels, stress response and cell cycle. The activity of these well conserved small polycations are also regulated by different mechanisms to assure their intracellular concentration and activity. Polyamine transporters have a major role in the regulation of the influx and efflux of the polyamines. Previously, two spermine family transporters SPBC409.08 and caf5 were characterized which were associated with cell size defects. The aim of this study is to better understand the role of caf5 plasma membrane spermine transporter and its functional interaction with spermidine transporters. Double mutants of caf5 and spermidine family transporters SPBC36.01c, SPBC36.03c and SPBC569.05c were formed in S. pombe. The double mutants were then put through growth rate analysis, meiotic progression and sporulation analysis, cell cycle analysis, and vulnerability to environmental stress. The results showed that the double mutants differentiate from each other in their combined mutation with caf5 in terms of cell size and DNA damage response, indicating redundant seperate pathways of spermine and spermidine transport in S. pombe.

Keywords:

S. pombe, Cell Size, Cell Cycle, Stress Response, Polyamines,

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[1] Katz AM, Tolokh IS, Pabit SA, Baker N, Onufriev AV, Pollack L. Spermine condenses DNA, but not RNA duplexes. Biophys J 2017; 112(1): 22–30.
[2] Fukuda W, Hidese R, Fujiwara S. Long-chain and branched polyamines in thermophilic microbes. In: Kusano T, Suzuki H editors. Polyamines: a universal molecular nexus for growth, survival, and specialized metabolism. Tokyo: Springer, 2015. pp. 15–26.
[3] Igarashi K, Kashiwagi K. Polyamine modulon in Escherichia coli: genes involved in the stimulation of cell growth by polyamines. J Biochem 2006; 139: 11–16.
[4] Uemura T, Higashi K, Takigawa M, Toida T, Kashiwagi K, Igarashi K. Polyamine modulon in yeast: stimulation of COX4 synthesis by spermidine at the level of translation. Int J Biochem Cell Biol 2009; 41: 2538–2545.
[5] Coyne HJ, Ciofi-Baffoni S, Banci L, Bertini I, Zhang L, George GN, Winge DR. The characterization and role of zinc bind- ing in yeast Cox4. J Biol Chem 2007; 282: 8926–8934.
[6] Örs Gevrekci A. The roles of polyamines in microorganisms. World J Microbiol Biotechnol 2018; 33(11): 204.
[7] Cohen SS. A Guide to the Polyamines. New York, NY, USA: Oxford University Press, 1998.
[8] Marina M, Maiale SJ, Rossi FR, Romero MF, Rivas EI, Garriz A, Ruiz OA, Pieckenstain FL. Apoplastic polyamine oxidation plays different roles in local responses of tobacco to infection by the necrotrophic fungus Sclerotinia sclerotiorum and the biotrophic bacterium Pseudomonas viridiflava. Plant Physiol 2008; 147: 2164–2178. [9] Valdés-Santiago L, Cervantes-Chávez JA, Ruiz-Herrera J. Ustilago maydis spermidine synthase is encoded by a chi- meric gene, required for morphogenesis, and indispensable for survival in the host. FEMS Yeast Res 2009; 9(6):923–935.
[10] Reigada C, Valera-Vera EA, Sayé M, Errasti AE, Avila CC, Miranda MR, Pereira CA. Trypanocidal effect of isotretinoin through the inhibition of polyamine and amino acid transporters in Trypanosoma cruzi. PloS Negl Trop Dis 2017; 11(3): e0005472.
[11] Wallace HM, Keir HM. Uptake and excretion of polyamines from baby hamster kidney cells (BHK-21/C13): the effect of serum on confluent cell cultures. Biochem Biophys Acta 1981; 676: 25-30.
[12] Wallace HM, Mackarel AJ. Regulation of polyamine acetylation and efflux in human cancer cells. Biochem Soc Trans 1998; 26: 571-575.
[13] Shah P, Nanduri B, Swiatlo E, Ma Y, Pendarvis K. Polyamine biosynthesis and transport mechanisms are crucial for fitness and pathogenesis of Streptococcus pneumoniae. Microbiology 2011; 157(2): 504–515.
[14] Jelsbak L, Thomsen LE, Wallrodt I, Jensen PR, Olsen JE. Polyamines are required for virulence in Salmonella enterica serovar Typhimurium. PLoS ONE 2012; 7(4): e36149.
[15] Örs Gevrekci A. The role of predicted spermidine family transporters in stress response and cell cycle in Schizosaccharomyces pombe. Turk J Biol 2017; 41: 419-427.
[16] Güngör İ, Örs Gevrekci A. The roles of SPBC409.08 and SPAC9.02c hypothetical genes in cell cycle and stress response, in Schizosaccharomyces pombe. Cell. Mol. Biol. 2016; 62 (4): 42-47.
[17] Moreno S, Klar A, Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods in Enzymology. 1991; 194: 795-823.
[18] Örs Gevrekci A. Functional characterization of spermine family transporter caf5+ in Schizosaccharomyces pombe (Lindner). Trakya Univ J Nat Sci, 2019; 20(2): xx-xx.
[19] Chattopadhyay MK, Tabor CW, Tabor H. Absolute requirement of spermidine for growth and cell cycle progression of fission yeast (Schizosaccharomyces pombe). Proc Natl Acad Sci USA 2002; 99(16): 10330–10334.
[20] Paulsen IT, Skurray RA. Topology, structure and evolution of two families of proteins involved in antibiotic and antiseptic resistance in eukaryotes and prokaryotes – an analysis. Gene.1993; 124(1): 1-11.
[21] Bäumgartner B, Möller H, Neumann T, Volkmer D. Preparation of thick silica coatings on carbon fibers with fine-structured silica nanotubes induced by a self-assembly process. Beilstein J Nanotechnol 2017; 8: 1145–1155.