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. 1998 Jun;10(6):981–993. doi: 10.1105/tpc.10.6.981

Pollen profilin function depends on interaction with proline-rich motifs.

B C Gibbon 1, L E Zonia 1, D R Kovar 1, P J Hussey 1, C J Staiger 1
PMCID: PMC144034  PMID: 9634586

Abstract

The actin binding protein profilin has dramatic effects on actin polymerization in vitro and in living cells. Plants have large multigene families encoding profilins, and many cells or tissues can express multiple profilin isoforms. Recently, we characterized several profilin isoforms from maize pollen for their ability to alter cytoarchitecture when microinjected into living plant cells and for their association with poly-L-proline and monomeric actin from maize pollen. In this study, we characterize a new profilin isoform from maize, which has been designated ZmPRO4, that is expressed predominantly in endosperm but is also found at low levels in all tissues examined, including mature and germinated pollen. The affinity of ZmPRO4 for monomeric actin, which was measured by two independent methods, is similar to that of the three profilin isoforms previously identified in pollen. In contrast, the affinity of ZmPRO4 for poly-L-proline is nearly twofold higher than that of native pollen profilin and the other recombinant profilin isoforms. When ZmPRO4 was microinjected into plant cells, the effect on actin-dependent nuclear position was significantly more rapid than that of another pollen profilin isoform, ZmPRO1. A gain-of-function mutant (ZmPRO1-Y6F) was created and found to enhance poly-L-proline binding activity and to disrupt cytoarchitecture as effectively as ZmPRO4. In this study, we demonstrate that profilin isoforms expressed in a single cell can have different effects on actin in living cells and that the poly-L-proline binding function of profilin may have important consequences for the regulation of actin cytoskeletal dynamics in plant cells.

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Selected References

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  1. Bedinger P. A., Edgerton M. D. Developmental staging of maize microspores reveals a transition in developing microspore proteins. Plant Physiol. 1990 Feb;92(2):474–479. doi: 10.1104/pp.92.2.474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bedinger P. The remarkable biology of pollen. Plant Cell. 1992 Aug;4(8):879–887. doi: 10.1105/tpc.4.8.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Björkegren-Sjögren C., Korenbaum E., Nordberg P., Lindberg U., Karlsson R. Isolation and characterization of two mutants of human profilin I that do not bind poly(L-proline). FEBS Lett. 1997 Dec 1;418(3):258–264. doi: 10.1016/s0014-5793(97)01376-8. [DOI] [PubMed] [Google Scholar]
  4. Björkegren C., Rozycki M., Schutt C. E., Lindberg U., Karlsson R. Mutagenesis of human profilin locates its poly(L-proline)-binding site to a hydrophobic patch of aromatic amino acids. FEBS Lett. 1993 Oct 25;333(1-2):123–126. doi: 10.1016/0014-5793(93)80388-b. [DOI] [PubMed] [Google Scholar]
  5. Bubb M. R., Baines I. C., Korn E. D. Localization of actobindin, profilin I, profilin II, and phosphatidylinositol-4,5-bisphosphate (PIP2) in Acanthamoeba castellanii. Cell Motil Cytoskeleton. 1998;39(2):134–146. doi: 10.1002/(SICI)1097-0169(1998)39:2<134::AID-CM4>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  6. Cao L. G., Babcock G. G., Rubenstein P. A., Wang Y. L. Effects of profilin and profilactin on actin structure and function in living cells. J Cell Biol. 1992 Jun;117(5):1023–1029. doi: 10.1083/jcb.117.5.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carlsson L., Nyström L. E., Sundkvist I., Markey F., Lindberg U. Actin polymerizability is influenced by profilin, a low molecular weight protein in non-muscle cells. J Mol Biol. 1977 Sep 25;115(3):465–483. doi: 10.1016/0022-2836(77)90166-8. [DOI] [PubMed] [Google Scholar]
  8. Christensen H. E., Ramachandran S., Tan C. T., Surana U., Dong C. H., Chua N. H. Arabidopsis profilins are functionally similar to yeast profilins: identification of a vascular bundle-specific profilin and a pollen-specific profilin. Plant J. 1996 Aug;10(2):269–279. doi: 10.1046/j.1365-313x.1996.10020269.x. [DOI] [PubMed] [Google Scholar]
  9. Clore A. M., Dannenhoffer J. M., Larkins B. A. EF-1[alpha] Is Associated with a Cytoskeletal Network Surrounding Protein Bodies in Maize Endosperm Cells. Plant Cell. 1996 Nov;8(11):2003–2014. doi: 10.1105/tpc.8.11.2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cooper J. A., Pollard T. D. Methods to measure actin polymerization. Methods Enzymol. 1982;85(Pt B):182–210. doi: 10.1016/0076-6879(82)85021-0. [DOI] [PubMed] [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Domke T., Federau T., Schlüter K., Giehl K., Valenta R., Schomburg D., Jockusch B. M. Birch pollen profilin: structural organization and interaction with poly-(L-proline) peptides as revealed by NMR. FEBS Lett. 1997 Jul 14;411(2-3):291–295. doi: 10.1016/s0014-5793(97)00719-9. [DOI] [PubMed] [Google Scholar]
  13. Evangelista M., Blundell K., Longtine M. S., Chow C. J., Adames N., Pringle J. R., Peter M., Boone C. Bni1p, a yeast formin linking cdc42p and the actin cytoskeleton during polarized morphogenesis. Science. 1997 Apr 4;276(5309):118–122. doi: 10.1126/science.276.5309.118. [DOI] [PubMed] [Google Scholar]
  14. Fedorov A. A., Ball T., Mahoney N. M., Valenta R., Almo S. C. The molecular basis for allergen cross-reactivity: crystal structure and IgE-epitope mapping of birch pollen profilin. Structure. 1997 Jan 15;5(1):33–45. doi: 10.1016/s0969-2126(97)00164-0. [DOI] [PubMed] [Google Scholar]
  15. Finkel T., Theriot J. A., Dise K. R., Tomaselli G. F., Goldschmidt-Clermont P. J. Dynamic actin structures stabilized by profilin. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1510–1514. doi: 10.1073/pnas.91.4.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Frazier J. A., Field C. M. Actin cytoskeleton: are FH proteins local organizers? Curr Biol. 1997 Jul 1;7(7):R414–R417. doi: 10.1016/s0960-9822(06)00205-3. [DOI] [PubMed] [Google Scholar]
  17. Gibbon B. C., Ren H., Staiger C. J. Characterization of maize (Zea mays) pollen profilin function in vitro and in live cells. Biochem J. 1997 Nov 1;327(Pt 3):909–915. doi: 10.1042/bj3270909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gieselmann R., Kwiatkowski D. J., Janmey P. A., Witke W. Distinct biochemical characteristics of the two human profilin isoforms. Eur J Biochem. 1995 May 1;229(3):621–628. doi: 10.1111/j.1432-1033.1995.tb20506.x. [DOI] [PubMed] [Google Scholar]
  19. Grote M., Vrtala S., Valenta R. Monitoring of two allergens, Bet v I and profilin, in dry and rehydrated birch pollen by immunogold electron microscopy and immunoblotting. J Histochem Cytochem. 1993 May;41(5):745–750. doi: 10.1177/41.5.8468456. [DOI] [PubMed] [Google Scholar]
  20. Haarer B. K., Petzold A. S., Brown S. S. Mutational analysis of yeast profilin. Mol Cell Biol. 1993 Dec;13(12):7864–7873. doi: 10.1128/mcb.13.12.7864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hartwig J. H., Kwiatkowski D. J. Actin-binding proteins. Curr Opin Cell Biol. 1991 Feb;3(1):87–97. doi: 10.1016/0955-0674(91)90170-4. [DOI] [PubMed] [Google Scholar]
  22. Hatano S. Actin-binding proteins in cell motility. Int Rev Cytol. 1994;156:199–273. doi: 10.1016/s0074-7696(08)62255-x. [DOI] [PubMed] [Google Scholar]
  23. Haugwitz M., Noegel A. A., Rieger D., Lottspeich F., Schleicher M. Dictyostelium discoideum contains two profilin isoforms that differ in structure and function. J Cell Sci. 1991 Nov;100(Pt 3):481–489. doi: 10.1242/jcs.100.3.481. [DOI] [PubMed] [Google Scholar]
  24. Hess M. W., Mittermann I., Luschnig C., Valenta R. Immunocytochemical localisation of actin and profilin in the generative cell of angiosperm pollen: TEM studies on high-pressure frozen and freeze-substituted Ledebouria socialis Roth (Hyacinthaceae). Histochem Cell Biol. 1995 Dec;104(6):443–451. doi: 10.1007/BF01464334. [DOI] [PubMed] [Google Scholar]
  25. Hightower R. C., Meagher R. B. The molecular evolution of actin. Genetics. 1986 Sep;114(1):315–332. doi: 10.1093/genetics/114.1.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Huang S., McDowell J. M., Weise M. J., Meagher R. B. The Arabidopsis profilin gene family. Evidence for an ancient split between constitutive and pollen-specific profilin genes. Plant Physiol. 1996 May;111(1):115–126. doi: 10.1104/pp.111.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hussey P. J., Haas N., Hunsperger J., Larkin J., Snustad D. P., Silflow C. D. The beta-tubulin gene family in Zea mays: two differentially expressed beta-tubulin genes. Plant Mol Biol. 1990 Dec;15(6):957–972. doi: 10.1007/BF00039438. [DOI] [PubMed] [Google Scholar]
  28. Imamura H., Tanaka K., Hihara T., Umikawa M., Kamei T., Takahashi K., Sasaki T., Takai Y. Bni1p and Bnr1p: downstream targets of the Rho family small G-proteins which interact with profilin and regulate actin cytoskeleton in Saccharomyces cerevisiae. EMBO J. 1997 May 15;16(10):2745–2755. doi: 10.1093/emboj/16.10.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Karakesisoglou I., Schleicher M., Gibbon B. C., Staiger C. J. Plant profilins rescue the aberrant phenotype of profilin-deficient Dictyostelium cells. Cell Motil Cytoskeleton. 1996;34(1):36–47. doi: 10.1002/(SICI)1097-0169(1996)34:1<36::AID-CM4>3.0.CO;2-G. [DOI] [PubMed] [Google Scholar]
  30. Lambrechts A., Verschelde J. L., Jonckheere V., Goethals M., Vandekerckhove J., Ampe C. The mammalian profilin isoforms display complementary affinities for PIP2 and proline-rich sequences. EMBO J. 1997 Feb 3;16(3):484–494. doi: 10.1093/emboj/16.3.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lin Y., Yang Z. Inhibition of Pollen Tube Elongation by Microinjected Anti-Rop1Ps Antibodies Suggests a Crucial Role for Rho-Type GTPases in the Control of Tip Growth. Plant Cell. 1997 Sep;9(9):1647–1659. doi: 10.1105/tpc.9.9.1647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Machesky L. M., Goldschmidt-Clermont P. J., Pollard T. D. The affinities of human platelet and Acanthamoeba profilin isoforms for polyphosphoinositides account for their relative abilities to inhibit phospholipase C. Cell Regul. 1990 Nov;1(12):937–950. doi: 10.1091/mbc.1.12.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mahoney N. M., Janmey P. A., Almo S. C. Structure of the profilin-poly-L-proline complex involved in morphogenesis and cytoskeletal regulation. Nat Struct Biol. 1997 Nov;4(11):953–960. doi: 10.1038/nsb1197-953. [DOI] [PubMed] [Google Scholar]
  34. Manseau L., Calley J., Phan H. Profilin is required for posterior patterning of the Drosophila oocyte. Development. 1996 Jul;122(7):2109–2116. doi: 10.1242/dev.122.7.2109. [DOI] [PubMed] [Google Scholar]
  35. Mascarenhas J. P. Molecular Mechanisms of Pollen Tube Growth and Differentiation. Plant Cell. 1993 Oct;5(10):1303–1314. doi: 10.1105/tpc.5.10.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Meagher R. B. Divergence and differential expression of actin gene families in higher plants. Int Rev Cytol. 1991;125:139–163. doi: 10.1016/s0074-7696(08)61218-8. [DOI] [PubMed] [Google Scholar]
  37. Metzler W. J., Bell A. J., Ernst E., Lavoie T. B., Mueller L. Identification of the poly-L-proline-binding site on human profilin. J Biol Chem. 1994 Feb 11;269(6):4620–4625. [PubMed] [Google Scholar]
  38. Pantaloni D., Carlier M. F. How profilin promotes actin filament assembly in the presence of thymosin beta 4. Cell. 1993 Dec 3;75(5):1007–1014. doi: 10.1016/0092-8674(93)90544-z. [DOI] [PubMed] [Google Scholar]
  39. Perelroizen I., Didry D., Christensen H., Chua N. H., Carlier M. F. Role of nucleotide exchange and hydrolysis in the function of profilin in action assembly. J Biol Chem. 1996 May 24;271(21):12302–12309. doi: 10.1074/jbc.271.21.12302. [DOI] [PubMed] [Google Scholar]
  40. Perelroizen I., Marchand J. B., Blanchoin L., Didry D., Carlier M. F. Interaction of profilin with G-actin and poly(L-proline). Biochemistry. 1994 Jul 19;33(28):8472–8478. doi: 10.1021/bi00194a011. [DOI] [PubMed] [Google Scholar]
  41. Petrella E. C., Machesky L. M., Kaiser D. A., Pollard T. D. Structural requirements and thermodynamics of the interaction of proline peptides with profilin. Biochemistry. 1996 Dec 24;35(51):16535–16543. doi: 10.1021/bi961498d. [DOI] [PubMed] [Google Scholar]
  42. Reinhard M., Giehl K., Abel K., Haffner C., Jarchau T., Hoppe V., Jockusch B. M., Walter U. The proline-rich focal adhesion and microfilament protein VASP is a ligand for profilins. EMBO J. 1995 Apr 18;14(8):1583–1589. doi: 10.1002/j.1460-2075.1995.tb07146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ren H., Gibbon B. C., Ashworth S. L., Sherman D. M., Yuan M., Staiger C. J. Actin Purified from Maize Pollen Functions in Living Plant Cells. Plant Cell. 1997 Aug;9(8):1445–1457. doi: 10.1105/tpc.9.8.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rothkegel M., Mayboroda O., Rohde M., Wucherpfennig C., Valenta R., Jockusch B. M. Plant and animal profilins are functionally equivalent and stabilize microfilaments in living animal cells. J Cell Sci. 1996 Jan;109(Pt 1):83–90. doi: 10.1242/jcs.109.1.83. [DOI] [PubMed] [Google Scholar]
  45. Schlüter K., Jockusch B. M., Rothkegel M. Profilins as regulators of actin dynamics. Biochim Biophys Acta. 1997 Nov 27;1359(2):97–109. doi: 10.1016/s0167-4889(97)00100-6. [DOI] [PubMed] [Google Scholar]
  46. Shah D. M., Hightower R. C., Meagher R. B. Genes encoding actin in higher plants: intron positions are highly conserved but the coding sequences are not. J Mol Appl Genet. 1983;2(1):111–126. [PubMed] [Google Scholar]
  47. Shen B., Carneiro N., Torres-Jerez I., Stevenson B., McCreery T., Helentjaris T., Baysdorfer C., Almira E., Ferl R. J., Habben J. E. Partial sequencing and mapping of clones from two maize cDNA libraries. Plant Mol Biol. 1994 Nov;26(4):1085–1101. doi: 10.1007/BF00040691. [DOI] [PubMed] [Google Scholar]
  48. Sheterline P., Clayton J., Sparrow J. Actin. Protein Profile. 1995;2(1):1–103. [PubMed] [Google Scholar]
  49. Sohn R. H., Chen J., Koblan K. S., Bray P. F., Goldschmidt-Clermont P. J. Localization of a binding site for phosphatidylinositol 4,5-bisphosphate on human profilin. J Biol Chem. 1995 Sep 8;270(36):21114–21120. doi: 10.1074/jbc.270.36.21114. [DOI] [PubMed] [Google Scholar]
  50. Staiger C. J., Goodbody K. C., Hussey P. J., Valenta R., Drøbak B. K., Lloyd C. W. The profilin multigene family of maize: differential expression of three isoforms. Plant J. 1993 Oct;4(4):631–641. doi: 10.1046/j.1365-313x.1993.04040631.x. [DOI] [PubMed] [Google Scholar]
  51. Staiger C. J., Yuan M., Valenta R., Shaw P. J., Warn R. M., Lloyd C. W. Microinjected profilin affects cytoplasmic streaming in plant cells by rapidly depolymerizing actin microfilaments. Curr Biol. 1994 Mar 1;4(3):215–219. doi: 10.1016/s0960-9822(00)00050-6. [DOI] [PubMed] [Google Scholar]
  52. Sun H. Q., Kwiatkowska K., Yin H. L. Actin monomer binding proteins. Curr Opin Cell Biol. 1995 Feb;7(1):102–110. doi: 10.1016/0955-0674(95)80051-4. [DOI] [PubMed] [Google Scholar]
  53. Taylor Loverine P., Hepler Peter K. POLLEN GERMINATION AND TUBE GROWTH. Annu Rev Plant Physiol Plant Mol Biol. 1997 Jun;48(NaN):461–491. doi: 10.1146/annurev.arplant.48.1.461. [DOI] [PubMed] [Google Scholar]
  54. Thorn K. S., Christensen H. E., Shigeta R., Huddler D., Shalaby L., Lindberg U., Chua N. H., Schutt C. E. The crystal structure of a major allergen from plants. Structure. 1997 Jan 15;5(1):19–32. doi: 10.1016/s0969-2126(97)00163-9. [DOI] [PubMed] [Google Scholar]
  55. Valenta R., Duchêne M., Pettenburger K., Sillaber C., Valent P., Bettelheim P., Breitenbach M., Rumpold H., Kraft D., Scheiner O. Identification of profilin as a novel pollen allergen; IgE autoreactivity in sensitized individuals. Science. 1991 Aug 2;253(5019):557–560. doi: 10.1126/science.1857985. [DOI] [PubMed] [Google Scholar]
  56. Vidali L., Hepler P. K. Characterization and localization of profilin in pollen grains and tubes of Lilium longiflorum. Cell Motil Cytoskeleton. 1997;36(4):323–338. doi: 10.1002/(SICI)1097-0169(1997)36:4<323::AID-CM3>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  57. Watanabe N., Madaule P., Reid T., Ishizaki T., Watanabe G., Kakizuka A., Saito Y., Nakao K., Jockusch B. M., Narumiya S. p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin. EMBO J. 1997 Jun 2;16(11):3044–3056. doi: 10.1093/emboj/16.11.3044. [DOI] [PMC free article] [PubMed] [Google Scholar]

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