The Characterization of Ligand-specific Maize (Zea Mays) Profilin Mutants

Overview

Journal Biochem J

Specialty Biochemistry

Date 2001 Aug 4

PMID 11485551

Citations 11

Authors

D R Kovar

B K Drobak

D A Collings

C J Staiger

Affiliations

Soon will be listed here.

Abstract

Profilins are low-molecular-mass (12-15 kDa) cytosolic proteins that are major regulators of actin assembly in all eukaryotic cells. In general, profilins from evolutionarily diverse organisms share the ability to bind to G-actin, poly-(L-proline) (PLP) and proline-rich proteins, and polyphosphoinositides. However, the functional importance of each of these interactions remains unclear and might differ between organisms. We investigated the importance of profilin's interaction with its various ligands in plant cells by characterizing four maize (Zea mays) profilin 5 (ZmPRO5) mutants that had single amino acid substitutions in the presumed sites of ligand interaction. Comparisons in vitro with wild-type ZmPRO5 showed that these mutations altered ligand association specifically. ZmPRO5-Y6F had a 3-fold increased affinity for PLP, ZmPRO5-Y6Q had a 5-fold decreased affinity for PLP, ZmPRO5-D8A had a 2-fold increased affinity for PtdIns(4,5)P(2) and ZmPRO5-K86A had a 35-fold decreased affinity for G-actin. When the profilins were microinjected into Tradescantia stamen hair cells, ZmPRO5-Y6F increased the rate of nuclear displacement in stamen hairs, whereas ZmPRO5-K86A decreased the rate. Mutants with a decreased affinity for PLP (ZmPRO5-Y6Q) or an enhanced affinity for PtdIns(4,5)P(2) (ZmPRO5-D8A) were not significantly different from wild-type ZmPRO5 in affecting nuclear position. These results indicate that plant profilin's association with G-actin is extremely important and further substantiate the simple model that profilin acts primarily as a G-actin-sequestering protein in plant cells. Furthermore, interaction with proline-rich binding partners might also contribute to regulating profilin's effect on actin assembly in plant cells.

Citing Articles

The role of the actin cytoskeleton in plant cell signaling.

Drobak B, Franklin-Tong V, Staiger C New Phytol. 2021; 163(1):13-30.

PMID: 33873778 DOI: 10.1111/j.1469-8137.2004.01076.x.

Structure and functions of profilins.

Krishnan K, Moens P Biophys Rev. 2017; 1(2):71-81.

PMID: 28509986 PMC: 5425664. DOI: 10.1007/s12551-009-0010-y.

Profilin-Dependent Nucleation and Assembly of Actin Filaments Controls Cell Elongation in Arabidopsis.

Cao L, Henty-Ridilla J, Blanchoin L, Staiger C Plant Physiol. 2015; 170(1):220-33.

PMID: 26574597 PMC: 4704583. DOI: 10.1104/pp.15.01321.

When fat is not bad: the regulation of actin dynamics by phospholipid signaling molecules.

Pleskot R, Pejchar P, Staiger C, Potocky M Front Plant Sci. 2014; 5:5.

PMID: 24478785 PMC: 3899574. DOI: 10.3389/fpls.2014.00005.

Analysis of the effects of polymorphism on pollen profilin structural functionality and the generation of conformational, T- and B-cell epitopes.

Jimenez-Lopez J, Rodriguez-Garcia M, Alche J PLoS One. 2013; 8(10):e76066.

PMID: 24146818 PMC: 3798325. DOI: 10.1371/journal.pone.0076066.

References

Pantaloni D, Carlier M . How profilin promotes actin filament assembly in the presence of thymosin beta 4. Cell. 1993; 75(5):1007-14. DOI: 10.1016/0092-8674(93)90544-z. View

Ballweber E, Giehl K, Hannappel E, Huff T, Jockusch B, Mannherz H . Plant profilin induces actin polymerization from actin : beta-thymosin complexes and competes directly with beta-thymosins and with negative co-operativity with DNase I for binding to actin. FEBS Lett. 1998; 425(2):251-5. DOI: 10.1016/s0014-5793(98)00240-3. View

Kang F, Purich D, Southwick F . Profilin promotes barbed-end actin filament assembly without lowering the critical concentration. J Biol Chem. 1999; 274(52):36963-72. DOI: 10.1074/jbc.274.52.36963. View

Witke W, Podtelejnikov A, Nardo A, Sutherland J, Gurniak C, Dotti C . In mouse brain profilin I and profilin II associate with regulators of the endocytic pathway and actin assembly. EMBO J. 1998; 17(4):967-76. PMC: 1170446. DOI: 10.1093/emboj/17.4.967. View

Petrella E, Machesky L, Kaiser D, Pollard T . Structural requirements and thermodynamics of the interaction of proline peptides with profilin. Biochemistry. 1996; 35(51):16535-43. DOI: 10.1021/bi961498d. View

Valster A, Pierson E, Valenta R, Hepler P, Emons A . Probing the Plant Actin Cytoskeleton during Cytokinesis and Interphase by Profilin Microinjection. Plant Cell. 2002; 9(10):1815-1824. PMC: 157024. DOI: 10.1105/tpc.9.10.1815. View

Goldschmidt-Clermont P, Machesky L, Baldassare J, Pollard T . The actin-binding protein profilin binds to PIP2 and inhibits its hydrolysis by phospholipase C. Science. 1990; 247(4950):1575-8. DOI: 10.1126/science.2157283. View

Jonckheere V, Lambrechts A, Vandekerckhove J, Ampe C . Dimerization of profilin II upon binding the (GP5)3 peptide from VASP overcomes the inhibition of actin nucleation by profilin II and thymosin beta4. FEBS Lett. 1999; 447(2-3):257-63. DOI: 10.1016/s0014-5793(99)00293-8. View

McGough A . F-actin-binding proteins. Curr Opin Struct Biol. 1998; 8(2):166-76. DOI: 10.1016/s0959-440x(98)80034-1. View

10.

Houk Jr T, Ue K . The measurement of actin concentration in solution: a comparison of methods. Anal Biochem. 1974; 62(1):66-74. DOI: 10.1016/0003-2697(74)90367-4. View

11.

Gibbon B, Zonia L, Kovar D, Hussey P, Staiger C . Pollen profilin function depends on interaction with proline-rich motifs. Plant Cell. 1998; 10(6):981-93. PMC: 144034. DOI: 10.1105/tpc.10.6.981. View

12.

Staiger C, Yuan M, Valenta R, Shaw P, Warn R, Lloyd C . Microinjected profilin affects cytoplasmic streaming in plant cells by rapidly depolymerizing actin microfilaments. Curr Biol. 1994; 4(3):215-9. DOI: 10.1016/s0960-9822(00)00050-6. View

13.

Goldschmidt-Clermont P, Furman M, Wachsstock D, Safer D, Nachmias V, Pollard T . The control of actin nucleotide exchange by thymosin beta 4 and profilin. A potential regulatory mechanism for actin polymerization in cells. Mol Biol Cell. 1992; 3(9):1015-24. PMC: 275662. DOI: 10.1091/mbc.3.9.1015. View

14.

Valenta R, Breiteneder H, Petternburger K, Breitenbach M, Rumpold H, Kraft D . Homology of the major birch-pollen allergen, Bet v I, with the major pollen allergens of alder, hazel, and hornbeam at the nucleic acid level as determined by cross-hybridization. J Allergy Clin Immunol. 1991; 87(3):677-82. DOI: 10.1016/0091-6749(91)90388-5. View

15.

Vidali L, Hepler P . Characterization and localization of profilin in pollen grains and tubes of Lilium longiflorum. Cell Motil Cytoskeleton. 1997; 36(4):323-38. DOI: 10.1002/(SICI)1097-0169(1997)36:4<323::AID-CM3>3.0.CO;2-6. View

16.

Cooley L, Verheyen E, Ayers K . chickadee encodes a profilin required for intercellular cytoplasm transport during Drosophila oogenesis. Cell. 1992; 69(1):173-84. DOI: 10.1016/0092-8674(92)90128-y. View

17.

Pollard T, Cooper J . Quantitative analysis of the effect of Acanthamoeba profilin on actin filament nucleation and elongation. Biochemistry. 1984; 23(26):6631-41. DOI: 10.1021/bi00321a054. View

18.

Pollard T, Almo S, Quirk S, Vinson V, Lattman E . Structure of actin binding proteins: insights about function at atomic resolution. Annu Rev Cell Biol. 1994; 10:207-49. DOI: 10.1146/annurev.cb.10.110194.001231. View

19.

Banno H, Chua N . Characterization of the arabidopsis formin-like protein AFH1 and its interacting protein. Plant Cell Physiol. 2000; 41(5):617-26. DOI: 10.1093/pcp/41.5.617. View

20.

Sohn R, Goldschmidt-Clermont P . Profilin: at the crossroads of signal transduction and the actin cytoskeleton. Bioessays. 1994; 16(7):465-72. DOI: 10.1002/bies.950160705. View