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Identification of Rice Kinesin 13A Interacting Proteins through Yeast Two-Hybrid Screening

Author Affiliations

  • 1 Department of Integrative Plant Science, Chung-Ang University, Anseong 456-756, KOREA
  • 2 National Academy of Agricultural Science, RDA, Suwon 441-707, KOREA

Int. Res. J. Biological Sci., Volume 2, Issue (8), Pages 20-26, August,10 (2013)


By using full-length rice kinesin 13A protein (OsKIN13A) as a bait construct and rice cDNA library as a prey, we obtained several interacting proteins through yeast two-hybrid (Y2H) screening. These interacting proteins were selected by beta-galactose filter assay after growing on SD-TLH media containing 25 mM of 3-aminotriazole (3-AT). All of selected interacting proteins were confirmed by sequencing and annotated according to the rice genome database, and grouped into seven functional categories, i.e. cellular processes, transport and binding protein, metabolic activity, signal transduction, molecular chaperone and other category. This study suggests that OsKIN13A protein is associated with wide range of biological processes such as development, signaling and immunity.


  1. Brady S.T., A novel ATPase from brain with properties expected for the fast axonal transport motor, Nature (Lond.),317, 7375 (1985)
  2. Vale R.D., Reese T.S. and Sheetz M. P., Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility, Cell,42(1), 3950 (1985)
  3. Asada T., Sonobe S. and Shibaoka H., Microtubule translocation in the cytokinetic apparatus of cultured tobacco cells, Nature.,350, 238241 (1991)
  4. Tiezzi A., Moscatelli A., Cai G., Bartalesi A. and CrestiM., An immunoreactive homolog of mammalian kinesin in Nicotiana tabacum pollen tubes, Cell Motil. Cytoskel.,21, 132137 (1992)
  5. Cai G., Bartalesi A., Del Casino C., Moscatelli A., Tiezzi A. and Cresti M., The kinesin-immunoreactive homologue from Nicotiana tabacum pollen tube: biochemical properties and subcellular localization, Planta,191, 496506 (1993)
  6. Lawrence C.J., Dawe, R.K., Christie K.R., Cleveland D.W., Dawson S.C., Endow S.A., Goldstein L.S.B., Goodson H.V., Hirokawa N., Howard J. et al., A standardized kinesin nomenclature, J. Cell Biol, 167,1922 (2004)
  7. Reddy A.S.N. and Day I.S., Kinesins in the Arabidopsisgenome: a comparative analysis among eukaryotes, BMC Genomics, 2 (2001)
  8. Zhong R., Burk D.H., Morrison W.H. and Ye Z.H., A kinesin-like protein is essential for oriented deposition of cellulose microfibrils and cell wall strength, Plant Cell, 14, 31013117 (2002)
  9. Lee Y.R., Li Y. and Liu B., Two Arabidopsisphragmoplast-associated kinesins play a critical role in cytokinesis during male gametogenesis, Plant Cell,19, 25952605 (2007)
  10. Ambrose J.C. and Cyr R., The kinesin ATK5 functions in early spindle assembly in Arabidopsis, Plant Cell,19, 226236 (2007)
  11. Yang X.Y., Chen Z.W., Xu T., Qu Z., Pan X.D., Qin X.H., Ren D.T. and Liu G.Q., Arabidopsis kinesin KP1 specifically interacts with VDAC3, a mitochondrial protein, and regulates respiration during seed germination at low temperature, Plant Cell,23, 10931106 (2011)
  12. Richardson D.N., Simmons M.P. and Reddy A.S., Comprehensive comparative analysis of kinesins in photosynthetic eukaryotes, BMC Genomics,, 18 (2006)
  13. Zhou S., Wang Y., Li W., Zhao Z., Ren Y., Wang Y., Gu S., Lin Q., Wang D., Jiang L., Su N., Zhang X., Liu L.,. Cheng Z, Lei C., Wang J., Guo X., Wu F., Ikehashi H., Wang H. and Wan J., Pollen semi-sterility1 encodes a kinesin-1-like protein important for male meiosis, anther dehiscence, and fertility in rice, Plant Cell,23, 111129 (2011)
  14. Zhang M., Zhang B., Qian Q., Yu Y., Li R., Zhang J., Liu X., Zeng D., Li J. and Zhou Y.. Brittle Culm 12, a dual-targeting kinesin-4 protein, controls cell-cycle progression and wall properties in rice, Plant J.,63, 312328 (2010)
  15. Li J., Jiang J., Qian Q., Xu Y., Zhang C., Xiao J., Du C., Luo W., Zou G., Chen M., Huang Y., Feng Y., Cheng Z., Yuan M. and Chong K.. Mutation of rice BC12/GDD1, which encodes a kinesin-like protein that binds to a GA biosynthesis gene promoter, leads to dwarfism with impaired cell elongation, Plant Cell,23, 628640 (2011)
  16. Fields S. and Song O., A novel genetic system to detect protein-protein interactions, Nature,340, 245246 (1989)
  17. Maithri S.K., Ramesh K.V., Mutangana D. and Sudha D, Molecular modeling and docking studies of PirB fusion protein from Photorhabdus Luminescens,Int. Res. J. Biological Sci.,1(8), 718 (2012)
  18. Martinez K.C., Teves F.G. and Ma. Reina Suzette B. Madamba M.R.S.B., Sequence analysis of putative luxS gene involved in prodigiosin biosynthesis from Philippine local strains of Serratia marcescens, Int. Res. J. Biological Sci.,2(4), 1319 (2013)
  19. Santhosh Kumar K., Kusuma L, Ramachandra N.B. and Mala N.V., Genetic variations among ecologically diverse species of Anurans at the level of Genus based on ISSR marker, Int. Res. J. Biological Sci., 1(7), 1119 (2012)
  20. Tania M., Runu C. and Utpal R., Study on enzyme-linked immunoassay and polymerase chain reaction for the identification and quantification of Staphylococcus aureus in pure culture and food samples, Int. Res. J. Biological Sci., 1(3), 3036 (2012)
  21. Balne N.G. and Ramachandra Rao C.S.V., Role of XmnIrestriction site polymorphism and JAK2 gene mutation in -Thalassemia, Int. Res. J. Biological Sci., 2(1), 4145 (2013)
  22. Gietz D., St Jean A., Woods R.A. and Schiestl R.H., Improved method for high effieciency transformation of intact yeast cells, Nucl. Acids Res.,20, 1425 (1992)
  23. Breeden L. and Nasmyth K., Regulation of the yeast HO gene, Cold Spring Harbor Symp. Quant. Biol.,50, 643650 (1995)
  24. Ling M., Merante F. and Robinson B.H., A rapid and reliable DNA preparation method for screening a large number of yeast clones by polymerase chain reaction, Nucl. Acid Res,23, 49244925 (1995)
  25. Leon O. and Roth M., Zinc fingers: DNA binding and protein–protein interactions, Biol. Res., 33, 2130 (2000)
  26. Wang J.Y., Xia X.L., Wang J.P. and Yin W.L., Stress responsive zinc-finger protein gene of Populus euphraticain tobacco enhances salt tolerance, J. Integr. Plant Biol,50(1), 5661 (2008)
  27. Lorick K.L., Jensen J.P., Fang S., Ong A.M., Hatakeyama S. and Weissman A.M., RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Proc. Natl. Acad. Sci. USA,96, 11364–11369 (1999)
  28. Joazeiro C.A. and Weissman A.M., RING finger proteins: mediators of ubiquitin ligase activity, Cell,102, 549552 (2000)
  29. Miernyk J.A., Duck N.B., Shatters Jr R.G. and Folk W.R., The 70-kilodalton heat shock cognate can act as a molecular chaperone during the membrane translocation of a plant secretory protein precursor, Plant Cell,4(7), 821829 (1992)
  30. Chamberlain L.H. and Burgoyne R.D., Activation of the ATPase activity of heat-shock protein Hsc70/Hsp70 by cysteine-string protein, Biochem. J.,322, 853858 (1997)
  31. Palavan-Unsal N., Buyuktuncer E. and Tufekci M.A., Programmed cell death in plants. J. Cell Mol. Biol.,, 923 (2005)
  32. Gilchrist D.G., Programmed cell death in plant disease: the purpose and promise of cellular suicide, Annu. Rev. Phytopathol.,36, 393414 (1998)
  33. Aalen R.B., Salehian Z. and Steinum T.M., Stability of barley aleurone transcripts: dependence on protein synthesis, influence of the starchy endosperm and destabilization by GA3, Physiol. Plant.,112(3), 403413 (2001)
  34. Huang Y.J., To K.Y., Yap M.N., Chiang W.J., Suen D.F. and Chen S.C.G., Cloning and characterization of leaf senescence up-regulated genes in sweet potato, Physiol. Plant.,113, 384391 (2001)
  35. Cui X., Fan B., Scholz J. and Chen Z., Roles of Arabidopsis cyclin-dependent kinase C complexes in cauliflower mosaic virus infection, plant growth, and development, Plant Cell,19(4), 13881402 (2007)
  36. Powell A.L., van Kan J., ten Have A., Visser J., et al.,. Transgenic expression of pear PGIP in tomato limits fungal colonization, Mol. Plant Microbe Interact., 13, 942950 (2000)
  37. Agüero C.B., Uratsu S.L., Greve C., Powell A.L.T., et al., Evaluation of tolerance to Pierce’s disease and Botrytis in transgenic plants of Vitis vinifera L. expressing the pear PGIP gene, Mol. Plant. Pathol., 4351 (2005)
  38. Ferrari S., Vairo D., Ausubel F.M., Cervone F. and De Lorenzo G., Tandemly duplicated Arabidopsis genes that encode polygalacturonase-inhibiting proteins are regulated coordinately by different signal transduction pathways in response to fungal infection, Plant Cell, 15, 93106 (2003)
  39. Manfredini C., Sicilia F., Ferrari S., Pontiggia D., Salvi G., Caprari C., Lorito M. and De Lorenzo G., Polygalacturonase-inhibiting protein 2 of Phaseolus vulgaris inhibits BcPG1, a polygalacturonase of Botrytis cinerea important for pathogenicity, and protects transgenic plants from infection, Physiol. Mol. Plant Pathol.,67, 108115 (2005)
  40. Hwang B.H., Bae H., Lim H.S., Kim K.B., Kim S.J., Im M.H., Park B.S., Kim D.S. and Kim J., Overexpression of polygalacturonase-inhibiting protein 2 (PGIP2) of Chinese cabbage (Brassica rapassp. pekinensis) increased resistance to the bacterial pathogen Pectobacterium carotovorum ssp. Carotovorum, Plant Cell Tiss. Org. Cult.,103, 293305 (2010)
  41. Devoto A., Leckie F., Lupotto E., Cervone F. and De Lorenzo G., The promoter of a gene encoding a polygalacturonase-inhibiting protein of Phaseolus vulgaris L. is activated by wounding but not by elicitors or pathogen infection, Planta,205, 165174 (1998)