Cytological and transcriptional analysis reveal phosphatidylinositol signaling pathway plays key role in mitotic division of <i>Pyropia yezoensis</i> -- Journal of Oceanology and Limnology,2022,40(3):1148-1159

	Cite this paper:
	Yunke ZHU, Xinran WANG, Bin SUN, Xianghai TANG, Yunxiang MAO. Cytological and transcriptional analysis reveal phosphatidylinositol signaling pathway plays key role in mitotic division of Pyropia yezoensis[J]. Journal of Oceanology and Limnology, 2022, 40(3): 1148-1159

Cytological and transcriptional analysis reveal phosphatidylinositol signaling pathway plays key role in mitotic division of Pyropia yezoensis

Yunke ZHU¹, Xinran WANG¹, Bin SUN¹, Xianghai TANG¹, Yunxiang MAO^2,3

1 Key Laboratory of Marine Genetics and Breeding(Ministry of Education), College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China;
2 MOE Key Laboratory of Utilization and Conservation for Tropical Marine Bioresources, College of Fisheries and Life Science, Hainan Tropical Ocean University, Sanya 572022, China;
3 Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266237, China

Abstract:

The phosphatidylinositol (PI) signaling system, a central regulator of eukaryotic metabolism, is widely found in eukaryotes for regulating a variety of cell activities. Most of the genes in the PI signaling system were found conserved in Pyropia yezoensis. In this experiment, wortmannin was used as an inhibitor to inhibit the activity of phosphatidylinositol-3 kinase (PI3K), an important regulator of the PI signaling system. After wortmannin treatment, the mitotic division of P. yezoensis was significantly inhibited in a dose-dependent manner, and the mitotic division percentage was reduced by 68.1% and 91.9% in the 5- and 10-μmol/L groups, respectively. When thalli were treated with wortmannin, the levels of reactive oxygen species (ROS) were significantly decreased. Furthermore, the expression level of PI3K was inhibited and the expression levels of downstream genes regulated by PI3K was significantly changed. In the PI3K-AGC signaling pathway, the expression levels of Serine/threonine protein kinase (AGC) and cyclindependent kinases A (CDKA) were downregulated, while WEE1 kinase gene (WEE1) was upregulated. Three nicotinamide adenine dinucleotide phosphate (NADPH) oxidase genes were downregulated after wortmannin treatment. These results indicate that the PI signaling system plays an important role in the regulation of cell activity in P. yezoensis. It was speculated that the growth and development of P. yezoensis might be regulated by P. yezoensis PI3K, which promoted the expression of the AGC gene and further regulates the expression of downstream WEE1 and CDKA genes to advance mitotic division, and also promoted the expression level of NADPH oxidase that regulates ROS homeostasis.

Key words: Pyropia yezoensis|phosphatidylinositol signaling system|reactive oxygen species|mitotic division

Received: 2021-04-23 Revised:

Tools

PDF (1573 KB) Free

Print this page

Add to favorites

Email this article to others

Authors

Articles by Yunke ZHU

Articles by Xinran WANG

Articles by Bin SUN

Articles by Xianghai TANG

Articles by Yunxiang MAO

References:
Alvarez B, Garrido E, Garcia-Sanz J A, Carrera A C.2003.Phosphoinositide 3-kinase activation regulates cell division time by coordinated control of cell mass and cell cycle progression rate.Journal of Biological Chemistry, 278(29):26466-26473, https://doi.org/10.1074/jbc.M300663200.
Aranda A, Sequedo L, Tolosa L, Quintas G, Burello E, Castell J V, Gombau L.2013.Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay:a quantitative method for oxidative stress assessment of nanoparticle-treated cells.Toxicology in Vitro, 27(2):954-963, https://doi.org/10.1016/j.tiv.2013.01.016.
Blouin N A, Brodie J A, Grossman A C, Xu P, Brawley S H.2011.Porphyra:a marine crop shaped by stress.Trends in Plant Science, 16(1):29-37, https://doi.org/10.1016/j.tplants.2010.10.004.
Campa C C, Martini M, De Santis M C, Hirsch E.2015.How PI3K-derived lipids control cell division.Frontiers in Cell and Developmental Biology, 3:61, https://doi.org/10.3389/fcell.2015.00061.
Cao H, Ni X L, Zhang C Y, Shi W S, Xu Y X, Yan Y M, Zhang F X.2017.Alterations in the proteome of wheat primary roots after wortmannin application during seed germination.Acta Physiologiae Plantarum, 39(10):223, https://doi.org/10.1007/s11738-017-2511-9.
Cauvin C, Echard A.2015.Phosphoinositides:lipids with informative heads and mastermind functions in cell division.Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1851(6):832-843, https://doi.org/10.1016/j.bbalip.2014.10.013.
Chen C J, Chen H, Zhang Y, Thomas H R, Frank M H, He Y H, Xia R.2020.TBtools:An integrative toolkit developed for interactive analyses of big biological data.Molecular Plant, 13(8):1194-1202, https://doi.org/10.1016/j.molp.2020.06.009.
Dangi S, Cha H, Shapiro P.2003.Requirement for phosphatidylinositol-3 kinase activity during progression through S-phase and entry into mitosis.Cellular Signalling, 15(7):667-675, https://doi.org/10.1016/S0898-6568(03)00002-0.
De Nadai C, Huitorel P, Chiri S, Ciapa B.1998.Effect of wortmannin, an inhibitor of phosphatidylinositol 3-kinase, on the first mitotic divisions of the fertilized sea urchin egg.Journal of Cell Science, 111(Pt 17):2507-2518, https://doi.org/10.1016/S1357-2725(98)00066-1.
Dieck C B, Boss W F, Perera I Y.2012.A role for phosphoinositides in regulating plant nuclear functions.Frontiers in Plant Science, 3:50, https://doi.org/10.3389/fpls.2012.00050.
Elge S, Brearley C, Xia H J, Kehr J L, Xue H W, MuellerRoeber B.2001.An Arabidopsis inositol phospholipid kinase strongly expressed in procambial cells:Synthesis of PtdIns(4,5)P₂ and PtdIns(3,4,5)P₃ in insect cells by 5-phosphorylation of precursors.The Plant Journal, 26(6):561-571, https://doi.org/10.1046/j.1365-313x.2001.01051.x.
Foreman J, Demidchik V, Bothwell J H F, Mylona P, Miedema H, Torres M A, Linstead P, Costa S, Brownlee C, Jones J D G, Davies J M, Dolan L.2003.Reactive oxygen species produced by NADPH oxidase regulate plant cell growth.Nature, 422(6930):442-446, https://doi.org/10.1038/nature01485.
Fujinami R, Yamada T, Nakajima A, Takagi S, Idogawa A, Kawakami E, Tsutsumi M, Imaichi R.2017.Root apical meristem diversity in extant lycophytes and implications for root origins.New Phytologist, 215(3):1210-1220, https://doi.org/10.1111/nph.14630.
Garg N, Manchanda G.2009.ROS generation in plants:boon or bane? Plant Biosystems:An International Journal Dealing with all Aspects of Plant Biology, 143(1):81-96, https://doi.org/10.1080/11263500802633626.
Hakeem K R, Rehman R U, Tahir I.2014.Plant Signaling:Understanding the Molecular Crosstalk.Springer, New Delhi.355p.
Heilmann I.2009.Using genetic tools to understand plant phosphoinositide signalling.Trends in Plant Science, 14(3):171-179, https://doi.org/10.1016/j.tplants.2008.12.002.
Heilmann I.2016.Phosphoinositide signaling in plant development.Development, 143(12):2044-2055, https://doi.org/10.1242/dev.136432.Joo J H, Yoo H J, Hwang I, Lee J S, Nam K H, Bae Y S.2005.Auxin-induced reactive oxygen species production requires the activation of phosphatidylinositol 3-kinase.FEBS Letters, 579(5):1243-1248, https://doi.org/10.1016/j.febslet.2005.01.018.
Katayama K, Fujita N, Tsuruo T.2005.Akt/Protein kinase B-dependent phosphorylation and Inactivation of WEE1Hu promote cell cycle progression at G₂/M transition.Molecular and Cellular Biology, 25(13):5725-5737, https://doi.org/10.1128/MCB.25.13.5725-5737.2005.
Kaur G, Pati P K.2016.Analysis of cis-acting Regulatory elements of respiratory burst oxidase homolog (Rboh) gene families in Arabidopsis and rice provides clues for their diverse functions.Computational Biology and Chemistry, 62:104-118, https://doi.org/10.1016/j.compbiolchem.2016.04.002.
Lee Y, Bak G, Choi Y, Chuang W I, Cho H T, Lee Y.2008.Roles of phosphatidylinositol 3-kinase in root hair growth.Plant Physiology, 147(2):624-635, https://doi.org/10.1104/pp.108.117341.
Lee Y, Munnik T, Lee Y.2010.Plant phosphatidylinositol 3-kinase.In:Munnik T ed.Lipid Signaling in Plants.Springer, Berlin.p.95-106.
Li L, Saga N, Mikami K.2008.Phosphatidylinositol 3-kinase activity and asymmetrical accumulation of F-actin are necessary for establishment of cell polarity in the early development of monospores from the marine red alga Porphyra yezoensis.Journal of Experimental Botany, 59(13):3575-3586, https://doi.org/10.1093/jxb/ern207.
Liu J, Zhou J, Xing D.2012.Phosphatidylinositol 3-kinase plays a vital role in regulation of rice seed vigor via altering NADPH oxidase activity.PLoS One, 7(3):e33817, https://doi.org/10.1371/journal.pone.0033817.
Livanos P, Apostolakos P, Galatis B.2012.Plant cell division:ROS homeostasis is required.Plant Signaling & Behavior, 7(7):771-778, https://doi.org/10.4161/psb.20530.
Marqués M, Kumar A, Poveda A M, Zuluaga S, Hernández C, Jackson S, Pasero P, Carrera A C, Verma I M.2009.Specific function of phosphoinositide 3-kinase β in the control of DNA replication.Proceedings of the National Academy of Sciences of the United States of America, 106(18):7525-7530, https://doi.org/10.1073/pnas.0812000106.
Maxwell K, Johnson G N.2000.Chlorophyll fluorescence-a practical guide.Journal of Experimental Botany, 51:659-668, https://doi.org/10.1093/jexbot/51.345.659.
Ni X L, Zhang F X.2014.PI3K is involved in nucleolar structure and function on root-tip meristematic cells of Triticum aestivum L.Acta Histochemica, 116(5):838-843, https://doi.org/10.1016/j.acthis.2014.02.004.
Powis G, Bonjouklian R, Berggren M M, Gallegos A, Abraham R, Ashendel C, Zalkow L, Matter W F, Dodge J, Grindey G.1994.Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3-kinase.Cancer Research, 54(9):2419-2423, https://doi.org/10.1007/s002620050075.
Ramanan R, Tran Q G, Cho D H, Jung J E, Kim B H, Shin S Y, Choi S H, Liu K H, Kim D S, Lee S J, Crespo J L, Lee H G, Oh H M, Kim H S.2018.The ancient phosphatidylinositol 3-kinase signaling system is a master regulator of energy and carbon metabolism in algae.Plant Physiology, 177(3):1050-1065, https://doi.org/10.1104/pp.17.01780.
Rastogi R P, Singh S P, Häder D P, Sinha R P.2010.Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2', 7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937.Biochemical and Biophysical Research Communications, 397(3):603-607, https://doi.org/10.1016/j.bbrc.2010.06.006.
Rushton P J, Somssich I E, Ringler P, Shen Q X J.2010.WRKY transcription factors.Trends in Plant Science, 15(5):247-258, https://doi.org/10.1016/j.tplants.2010.02.006.
Sato-Izawa K, Nakaba S, Tamura K, Yamagishi Y, Nakano Y, Nishikubo N, Kawai S, Kajita S, Ashikari M, Funada R, Katayama Y, Kitano H.2012.DWARF50 (D50), a rice(Oryza sativa L.) gene encoding inositol polyphosphate 5-phosphatase, is required for proper development of intercalary meristem.Plant, Cell & Environment, 35(11):2031-2044, https://doi.org/10.1111/j.1365-3040.2012.02534.x.
Seo M, Hanada A, Kuwahara A, Endo A, Okamoto M, Yamauchi Y, North H, Marion-Poll A, Sun T P, Koshiba T, Kamiya Y, Yamaguchi S, Nambara E.2006.Regulation of hormone metabolism in Arabidopsis seeds:phytochrome regulation of abscisic acid metabolism and abscisic acid regulation of gibberellin metabolism.The Plant Journal, 48(3):354-366, https://doi.org/10.1111/j.1365-313X.2006.02881.x.
Shigaki T, Bhattacharyya M K.2002.Nutrients induce an increase in inositol 1,4,5-trisphosphate in soybean cells:implication for the involvement of phosphoinositidespecific phospholipase C in DNA synthesis.Plant Biology, 4(1):53-61, https://doi.org/10.1055/s-2002-20436.
Snider C E, Willet A H, Chen J S, Arpağ G, Zanic M, Gould K L.2017.Phosphoinositide-mediated ring anchoring resists perpendicular forces to promote medial cytokinesis.The Journal of Cell Biology, 216(10):3041-3050, https://doi.org/10.1083/jcb.201705070.
Tang L, Qiu L P, Liu C, Du G Y, Mo Z L, Tang X H, Mao Y X.2019.Transcriptomic insights into innate immunity responding to red rot disease in red alga Pyropia yezoensis.International Journal of Molecular Sciences, 20(23):5970, https://doi.org/10.3390/ijms20235970.
Wang D M, Yu X Z, Xu K P, Bi G Q, Cao M, Zelzion E, Fu C X, Sun P P, Liu Y, Kong F N, Du G Y, Tang X H, Yang R J, Wang J H, Tang L, Wang L, Zhao Y J, Ge Y, Zhuang Y Y, Mo Z L, Chen Y, Gao T, Guan X W, Chen R, Qu W H, Sun B, Bhattacharya D, Mao Y X.2020.Pyropia yezoensis genome reveals diverse mechanisms of carbon acquisition in the intertidal environment.Nature Communications, 11(1):4028, https://doi.org/10.1038/s41467-020-17689-1.
Wang Y, Chu Y J, Xue H W.2012.Inositol polyphosphate 5-phosphatase-controlled Ins (1,4,5)P₃/Ca²⁺ is crucial for maintaining pollen dormancy and regulating early germination of pollen.Development, 139(12):2221-2233, https://doi.org/10.1242/dev.081224.
Wu S F, Wang S Z, Gao F, Li L Y, Zheng S Y, Yung W K A, Koul D.2018.Activation of WEE1 confers resistance to PI3K inhibition in Glioblastoma.Neuro-Oncology, 20(1):78-91, https://doi.org/10.1093/neuonc/nox128.
Xu X J, Liu X Y, Zhang Y.2018.Osthole inhibits gastric cancer cell proliferation through regulation of PI3K/AKT.PLoS One, 13(3):e0193449, https://doi.org/10.1371/journal.pone.0193449.
Yamada M, Han X W, Benfey P N.2020.RGF1 controls root meristem size through ROS signalling.Nature, 577(7788):85-88, https://doi.org/10.1038/s41586-019-1819-6.