拟南芥AtJ3与PKS5相互作用参与植物ABA响应

doi:10.7525/j.issn.1673-5102.2016.01.015

摘要/Abstract

摘要： 脱落酸(abscisic acid，ABA)在植物生长、发育及环境胁迫响应中有着广泛的作用。前期研究已鉴定了诸多参与植物ABA信号转导的元件。本研究以拟南芥(Arabidopsis thaliana)蛋白激酶PKS5(SOS2-like protein kinase 5)为诱饵蛋白，使用酵母双杂交筛选到与PKS5相互作用的蛋白分子伴侣AtJ3(Arabidopsis thaliana DnaJ homolog 3)。AtJ3 T-DNA突变体atj3-1与atj3-2表现出萌发期ABA表型。在外源ABA处理下，atj3-1与atj3-2种子发芽率降彽、幼苗黄化、生长矮小。atj3-1与PKS5双突变体atj3-1pks5-1、atj3-1pks5-3和atj3-1pks5-4表现出与AtJ3或PKS5突变体相同的ABA表型。亚细胞定位与转基因研究显示AtJ3与PKS5有相同的表达模式。免疫共沉淀与体外磷酸化测试确认AtJ3与PKS5存在相互作用并抑制PKS5激酶活性。研究结果表明：AtJ3与PKS5相互作用，通过抑制PKS5激酶活性共同参与植物ABA信号响应过程。

关键词: ABA响应, AtJ3, 相互作用, 磷酸化活性抑制, PKS5

Abstract: The phytohormone abscisic acid(ABA) has a wide range of important roles in plant growth and development as well as abiotic stress response. Previous studies identified the enormous components involved in ABA responses in plants. We identified a chaperon AtJ3(Arabidopsis thaliana DnaJ homolog 3; heat shock protein 40-like) using yeast two-hybrid assays in which PKS5 was used as bait. The AtJ3 T-DNA mutants atj3-1 and atj3-2 displayed ABA phenotypes. Seed germinations of atj3-1 and atj3-2 decreased, and the seedlings of them showed stunted growth and leaf chlorotic symptoms under ABA. Double mutants of atj3-1pks5-1, atj3-1pks5-3 and atj3-1pks5-4 had similar ABA phenotypes as mutants of AtJ3 or PKS5. Moreover, the assays of subcellular location and transgenic plants showed that AtJ3 has overlapping expression pattern with PKS5. By co-immunoprecipitation and in vitro phosphorylation, AtJ3 physically interacts with PKS5 and represses the PKS5 kinase activity. Therefore, AtJ3 interacts with PKS5 in response to ABA by repressing the PKS5 kinase activity.

Key words: ABA response, AtJ3, interaction, phosphorylation repress, PKS5

赵菲佚；焦成瑾；陈荃；贾贞；王太术；周辉. 拟南芥AtJ3与PKS5相互作用参与植物ABA响应[J]. 植物研究, 2016, 36(1): 105-115.

ZHAO Fei-Yi；JIAO Cheng-Jin；CHEN Quan；JIA Zhen；WANG Tai-Shu；ZHOU Hui. AtJ3 is Involved in ABA Response through Interaction with the PKS5 Kinase in Arabidopsis[J]. Bulletin of Botanical Research, 2016, 36(1): 105-115.

参考文献

1.Koornneef M,Leon-Kloosterziel K,Schwartz S H,et al.The genetic and molecular dissection of abscisic acid biosynthesis and signal transduction in Arabidopsis［J］.Plant Physiol Biochem,1998,36:83-89.

2.Leung J,Giraudat J.Abscisic acid signal transduction［J］.Annu Rev Plant Physiol Plant Mol Biol,1998,49:199-222.

3.McCourt P.Genetic analysis of hormone signaling［J］.Annu Rev Plant Physiol Plant Mol Biol,1999,50:219-243.

4.Shinozaki K,Yamaguchi S K.Molecular responses to dehydration and low temperature:Differences and cross-talk between two stress signaling pathways［J］.Curr Opin Plant Biol,2000,3:217-223.

5.Hrabak E M,Chan C W,Gribskov M,et al.The Arabidopsis CDPK-SnRK superfamily of protein kinases［J］.Plant Physiology,2003,132:666-680.

6.Weinl S,Kudla J.The CBL-CIPK Ca²⁺-decoding signaling network:function and perspectives［J］.New Phytologist,2009,184:517-528.

7.Jen S,Li Z,Jang Z,et al.Sugars as signaling molecules［J］.Curr Opin Plant Biol,1999,2:410-418.

8.Gong D M,Zhang C Q,Chen X Y,et al.Constitutive activation and transgenic evaluation of the function of an Arabidopsis PKS protein kinase［J］.J Bio Chem,2002,227:42088-42096.

9.Cheong Y H,Kim K N,Pandey G K,et al.CBL1,a calcium sensor that differentially regulates salt,drought,and cold responses in Arabidopsis［J］.Plant Cell,2003,15:1833-1845.

10.Qin Y Z,Guo M,Li X,et al.Stress responsive gene CIPK14 is involved in phytochrome A-mediated far-red light inhibition of greening in Arabidopsis［J］.Science China Life Sciences,2010,40:970-977.

11.Qin Y Z,Li X,Guo M,et al.Regulation of salt and ABA responses by CIPK14,a calcium sensor interacting protein kinase in Arabidopsis［J］.Science in China Series C Life Sciences,2008,38:446-457.

12.Fuglsang A T,Guo Y,Cuin T A,et al.Arabidopsis protein kinase PKS5 inhibits the plasma membrane H⁺-ATPase by preventing interaction with 14-3-3 protein［J］.Plant Cell,2007,19:1617-1634.

13.Xie C G,Lin H X,Deng X W,et al.Roles of SCaBP8 in salt stress response［J］.Plant Signaling & Behavior,2009,4:956-958.

14.Xie C,Zhou X,Deng X,Guo Y.PKS5,a SNF1-related kinase,interacts with and phosphorylates NPR1,and modulates expression of WRKY38 and WRKY62［J］.J Genet Genomics,2010,37:359-369.

15.Boston R S,Viitanen P V,Vierling E.Molecular chaperones and protein folding in plants［J］.Plant Mol Biol,1996,32:191-222.

16.Waters E R,Lee G J,Vierling E.Evolution,structure and function of the small heat shock proteins in plants［J］.J Exp Bot,1996,47:325-338.

17.Wang W,Vinocur B,Shoseyov O,et al.Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response［J］.Trends Plant Sci,2004,9:244-252.

18.Caplan A J,Cyr D M,Douglas M G.Eukaryotic homologues of Escherichia coli dnaJ:A diverse protein family that functions with hsp70 stress proteins［J］.Mol Biol Cell,1993,4:555-563.

19.Silver P A,Way J C.Eukaryotic DnaJ homologs and the specificity of Hsp70 activity［J］.Cell,1993,74:5-6.

20.Qian Y Q,Patel D,Hartl F U,et al.Nuclear magnetic resonance solution structure of the human Hsp40(HDJ-1) J-domain［J］.J Mol Biol,1996,260:224-235.

21.Qiu X B,Shao Y M,Miao S,et al.The diversity of the DnaJ/Hsp40 family,the crucial partners for Hsp70 chaperones［J］.Cell Mol Life Sci,2006,63:2560-2570.

22.Miernyk J A.The J-domain proteins of Arabidopsis thaliana:An unexpectedly large and diverse family of chaperones［J］.Cell Stress Chaperones,2001,6:209-218.

23.Shen L,Yu H.J3 regulation of flowering time is mainly contributed by its activity in leaves［J］.Plant Signal Behav,2011,6:601-603.

24.Shen L,Kang Y G,Liu L,et al.The J-domain protein J3 mediates the integration of flowering signals in Arabidopsis［J］.Plant Cell,2011,23:499-514.

25.Yang Y Q,Qin Y X,Xie C G,et al.The Arabidopsis chaperone J3 regulates the plasma membrane H⁺-ATPase through interaction with the PKS5 kinase［J］.Plant cell,2010,22:1313-1332.

26.Jen S.Signal transduction in maize and Arabidopsis mesophyll protoplasts［J］.Plant Physiol,2001,127:1466-1475.

27.Quan R,Lin H,Mendoza I,et al.SCABP8/CBL10,a putative calcium sensor,interacts with the protein kinase SOS2 to protect Arabidopsis shoots from salt stress［J］.Plant Cell,2007,19:1415-1431.

28.Guo Y,Halfter U,Ishitani M,et al.Molecular characterization of functional domains in the protein kinase SOS2 that is required for plant salt tolerance［J］.Plant Cell,2001,13:1383-1399.

29.Gong D,Guo Y,Schumaker K S,et al.The SOS3 family of calcium sensors and SOS2 family of protein kinases in Arabidopsis［J］.Plant Physiol,2004,134:919-926.

30.Guo Y,Xiong L,Song C P,et al.A calcium sensor and its interacting protein kinase are global regulators of abscisic acid signaling in Arabidopsis［J］.Dev Cell,2002,3:233-244.

31.Halfter U,Ishitani M,Zhu J K.The Arabidopsis SOS2 protein kinase physically interacts with and is activated by the calciumbinding protein SOS3［J］.Proc Natl Acad Sci USA,2000,97:3735-3740.

32.Quintero F J,Ohta M,Shi H,et al.Reconstitution in yeast of the Arabidopsis SOS signaling pathway for Na⁺ homeostasis［J］.Proc Natl Acad Sci USA,2002,99:9061-9066.

33.Lin H,Yang Y,Quan R,et al.Phosphorylation of SOS3-LIKE CALCIUM BINDING PROTEIN8 by SOS2 protein kinase stabilizes their protein complex and regulates salt tolerance in Arabidopsis［J］.Plant Cell,2009,21:1607-1619.

34.Xu J,Li H D,Chen L Q,et al.A protein kinase,interacting with two calcineurin B-like proteins,regulates K⁺ transporter AKT1 in Arabidopsis［J］.Cell,2006,125:1347-1360.

[1]	陈家兴, 王姝, 陈林丽, 侯夏丽, 杨庆祝, 尹任娅. 干旱条件对鬼针草和醉鱼草种间相互作用及生长的影响[J]. 植物研究, 2023, 43(5): 720-728.
[2]	令狐克念, 王姝. 不同生长阶段苘麻生物量分配对种群密度和土壤水分的响应[J]. 植物研究, 2023, 43(2): 272-280.
[3]	郭鲲1；刘瑞鹏1；张玲1,2；毛子军1*. 原始阔叶红松林下红松、蒙古栎混合凋落叶分解特征及相互作用研究[J]. 植物研究, 2015, 35(5): 716-723.