Abstract
Purple acid phosphatases (PAPs) are metallo-phosphoesterases. Their expression and function have not been systematically investigated in higher plants. In this work, we compared the transcript levels of 28 Arabidopsis PAP (AtPAP) genes in five Arabidopsis organs. The 28 members, although differed in their expression patterns in vegetative organs, were all transcribed in flower. Furthermore, the transcription of seven members (AtPAPs 6, 11, 14, 19, 23, 24 and 25) occurred predominantly in the flower. To begin dissecting the role of AtPAP genes in flower development, further expression and functional analyses were conducted using AtPAP23. Histochemical staining of transgenic plants expressing AtPAP23 promoter-beta-glucuronidase (GUS) gene construct revealed that AtPAP23 transcription was strong in flower apical meristems, but became restricted to petals and anther filaments in fully developed flower. A GST (glutathione S-transferase) fusion protein of AtPAP23 (GST:AtPAP23) was expressed in bacterial cells, and was found to contain significant amounts of Fe and Mn (whereas the control GST protein contained none). In biochemical tests, GST:AtPAP23 showed typical acid phosphatase activities. The fusion protein was also highly active on phosphoserine, but not phosphotyrosine. Despite its highly specific expression pattern and the demonstrated biochemical function of its protein product, the RNAi (RNA interference), T-DNA knock-out and overexpression lines of AtPAP23 were indistinguishable from wild type plants in the development of flower (or other organs). Interestingly, the Fe and Mn contents were found significantly increased in AtPAP23 overexpression lines, which may offer a new direction for further functional studies of AtPAPs in Arabidopsis.
Similar content being viewed by others
Abbreviations
- AGI:
-
Arabidopsis genome initiative
- AtPAP:
-
Arabidopsis thaliana purple acid phosphatase
- FAM:
-
flower apical meristem
- GST:
-
Glutathione S-transferase
- GUS:
-
β-glucuronidase
- ICP-MS:
-
inductively coupled plasma-mass spectroscopy
- IPTG:
-
isopropylthio-β-galactoside
- PAP:
-
Purple acid phosphatase
- PMSF:
-
phenyl-methylsulfonyl fluoride
- PNPP:
-
p-nitrophenyl phosphate
- SAM:
-
shoot apical meristem
- TRAP:
-
tartrate-resistant acid phosphatase
References
O. Alcantara S.V. Reddy G.D. Roodman D.H. Boldt (1994) ArticleTitleTranscriptional regulation of the tartrate-resistant acid phosphatase (TRAP) gene by iron Biochem. J. 298 421–425 Occurrence Handle8135751
D. Barford A.K. Das M.P. Egloff (1998) ArticleTitleThe structure and mechanism of protein phosphatases: insights into catalysis and regulation Annu. Rev. Biomol. Struct. 27 133–164 Occurrence Handle10.1146/annurev.biophys.27.1.133
G.A. Baumbach C.M. Ketcham D.E. Richardson F.W. Bazer R.M. Roberts (1986) ArticleTitleIsolation and characterization of a high molecular weight stable pink form of uteroferrin from uterine secretions and allantoic fluid of pigs J. Biol. Chem. 261 12869–12878 Occurrence Handle3017991
A.A. Baykov O.A. Evtushenko S.M. Avaeva (1988) ArticleTitleA malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay Anal. Biochem. 171 266–270 Occurrence Handle10.1016/0003-2697(88)90484-8 Occurrence Handle3044186
N. Bechtold J. Ellis G. Pelletier (1993) ArticleTitleIn plantaAgrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants C.R. Acad. Sci. 316 1194–1199
N. Bouche D. Bouchez (2001) ArticleTitleArabidopsis gene knockout: phenotypes wanted Curr. Opin. Plant Biol. 4 111–117 Occurrence Handle10.1016/S1369-5266(00)00145-X Occurrence Handle11228432
G.G. Bozzo K.G. Raghothama W.C. Plaxton (2002) ArticleTitlePurification and characterization of two secreted purple acid phosphatase isozymes from phosphate-starved tomato (Lycopersicon esculentum) cell cultures Eur. J. Biochem. 269 6278–6286 Occurrence Handle10.1046/j.1432-1033.2002.03347.x Occurrence Handle12473124
G.G. Bozzo K.G. Raghothama W.C. Plaxton (2004) ArticleTitleStructural and kinetic properties of a novel purple acid phosphatase from phosphate-starved tomato (Lycopersicon esculentum) cell cultures Biochem. J. 377 419–428 Occurrence Handle10.1042/BJ20030947 Occurrence Handle14521509
A.G. Cashikar R. Kumaresan N.M. Rao (1997) ArticleTitleBiochemical characterization and subcellular localization of the red kidney bean purple acid phosphatase Plant Physiol. 114 907–915 Occurrence Handle12223752
J.C. Pozo Particledel I. Allona V. Rubio A. Leyva A. la Pena Particlede C. Aragoncillo J. Paz-Ares (1999) ArticleTitleA type 5 acid phosphatase gene from Arabidopsis thaliana is induced by phosphate starvation and by some other types of phosphate mobilizing/oxidative stress conditions Plant J. 19 579–589 Occurrence Handle10.1046/j.1365-313X.1999.00562.x Occurrence Handle10504579
K. Doi B.C. Antanaitis P. Aisen (1988) ArticleTitleThe binuclear iron centers of uteroferrin and the purple acid phosphatases Struct. Bond. 70 1–26
D.P. Dixon M. Lith Particlevan R. Edwards A. Benham (2003) ArticleTitleCloning and initial characterization of the Arabidopsis thaliana endoplasmic reticulum oxidoreductins Antioxid Redox Signal 5 389–396 Occurrence Handle10.1089/152308603768295122 Occurrence Handle13678526
A. Durmus C. Eichen B.H. Sift A. Kratel R. Kappl J. Hüttermann B. Krebs (1999) ArticleTitleThe active site of purple acid phosphatase from sweet potatoes (Ipomoea batatas) Eur. J. Biochem. 260 709–716 Occurrence Handle10.1046/j.1432-1327.1999.00230.x Occurrence Handle10102999
B. EK-Rylander M. Flores M. Wendel D. Heinegard G. Andersson (1994) ArticleTitleDephosphorylation of osteopontin and bone sialoprotein by osteoclastic tartrate-resistant acid phosphatase. Modulation of osteoclast adhesion in vitro J.␣Biol. Chem. 269 14853–14856 Occurrence Handle8195113
C. Ferrándiz Q. Gu R. Martienssen M.F. Yanofsky (2000) ArticleTitleRedundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1 and CAULIFLOWER Development 127 725–734 Occurrence Handle10648231
V. Garcia M. Salanoubat N. Choisne A. Tissier (2000) ArticleTitleAn ATM homologue from Arabidopsis thaliana: complete genomic organization and expression analysis Nucleic Acids Res. 28 1692–1699 Occurrence Handle10.1093/nar/28.8.1692 Occurrence Handle10734187
K.L. Guan J.E. Dixon (1991) ArticleTitleEukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione-S-transferase Anal. Biochem. 192 262–267 Occurrence Handle10.1016/0003-2697(91)90534-Z Occurrence Handle1852137
F. Guerineau A. Lucy P. Mullineaux (1992) ArticleTitleEffect of two consensus sequences preceding the translation initiator codon on gene expression in plant protoplasts Plant Mol. Biol. 18 815–818 Occurrence Handle10.1007/BF00020027 Occurrence Handle1373083
S. Haran S. Logendra M. Seskar M. Bratanova I. Raskin (2000) ArticleTitleCharacterization of Arabidopsis acid phosphatase promoter and regulation of acid phosphatase expression Plant Physiol. 124 615–626 Occurrence Handle10.1104/pp.124.2.615 Occurrence Handle11027712
A.R. Hayman T.M. Cox (1994) ArticleTitlePurple acid phosphatase of the human macrophage and osteoclast J. Biol. Chem. 269 1294–1300 Occurrence Handle8288593
A.R. Hayman S.J. Jones A. Boyde D. Foster W. Colledge M.B. Carlton M.J. Evans T.M. Cox (1996) ArticleTitleMice lacking tartrate-resistant acid phosphates (Acp5) have disrupted endochondral ossification and mild osteopetrosis Development 122 3151–3162 Occurrence Handle8898228
C.E. Hegeman E.A. Grabau (2001) ArticleTitleA novel phytase with similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings Plant Physiol. 126 1598–1608 Occurrence Handle10.1104/pp.126.4.1598 Occurrence Handle11500558
A. Herbik C. Bolling T.J. Buckhout (2002) ArticleTitleThe involvement of a multicopper oxidase in iron uptake by the green algae Chlamydomonas reinhardtii Plant Physiol. 130 2039–2048 Occurrence Handle10.1104/pp.013060 Occurrence Handle12481087
D. Honys D. Twell (2003) ArticleTitleComparative analysis of Arabidopsis pollen transcriptome Plant Physiol. 132 640–652 Occurrence Handle10.1104/pp.103.020925 Occurrence Handle12805594
Y. Hu F. Bao J. Li (2000) ArticleTitlePromotive effect of brassinosteroids on cell division involves a distinct CycD3-induction pathway in Arabidopsis Plant J. 24 693–701 Occurrence Handle10.1046/j.1365-313x.2000.00915.x Occurrence Handle11123807
L. Hunt L. Otterhag J.C. Lee T. Lasheen J. Hunt M. Seki K Shinozaki M. Sommarin D.J. Gilmour C. Pical J.E. Gray (2004) ArticleTitleGene-specific expression and calcium activation of Arabidopsis thaliana phospholipase C isoforms New Phytol. 162 643–654 Occurrence Handle10.1111/j.1469-8137.2004.01069.x
R.A. Jefferson (1987) ArticleTitleAssaying chimeric genes in plants: the GUS gene fusion system Plant Mol. Biol. Rep. 5 387–405
R. Kaida K. Sage-Ono H. Kamada H. Okuyama K. Syono T.S. Kaneko (2003) ArticleTitleIsolation and characterization of four cell wall purple acid phosphatase genes from tobacco cells Biochem. Biophys. Acta 1625 134–140 Occurrence Handle12531472
S.R. Kim A. Gynheung (1996) ArticleTitleIsolation and characterization of a pollen-specific cDNA clone from Easter lily J.␣Plant Biol. 39 197–202
T. Klabunde B. Krebs (1997) ArticleTitleThe dimetal center in purple acid phosphatases Struct. Bond. 89 177–198
M. Kusaba K. Dwyer J. Hendershot J. Vrebalov J.B. Nasrallah M.E. Nasrallah (2001) ArticleTitleSelf-incompatibility in the genus Arabidopsis: characterization of the S locus in the outcrossing A. lyrata and its autogamous relative A. thaliana Plant Cell 13 627–643 Occurrence Handle10.1105/tpc.13.3.627 Occurrence Handle11251101
T. Kusudo T. Sakaki K. Inouye (2003) ArticleTitlePurification and characterization of purple acid phosphatase PAP1 from dry powder of sweet potato Biosci. Biotechnol. Biochem. 67 1609–1611 Occurrence Handle10.1271/bbb.67.1609 Occurrence Handle12913313
D. Li H. Zhu K. Liu X. Liu G. Leggewie M. Udvardi D. Wang (2002) ArticleTitlePurple acid phosphatases of Arabidopsis thaliana: comparative analysis and differential regulation by phosphate deprivation J. Biol. Chem. 277 27772–27781 Occurrence Handle10.1074/jbc.M204183200 Occurrence Handle12021284
G. Li K. Liu S.A. Baldwin D. Wang (2003) ArticleTitleEquilibrative nucleoside transporters of Arabidopsis thaliana: cDNA cloning, expression pattern, and analysis of transport activities J. Biol. Chem. 278 35732–35742 Occurrence Handle10.1074/jbc.M304768200 Occurrence Handle12810710
P. Ling R.M. Roberts (1993) ArticleTitleUteroferrin and intracellular tartrate-resistant acid phosphatases are the products of the same gene J. Biol. Chem. 268 6896–6902 Occurrence Handle8463220
L. Ma N. Sun X. Liu Y. Jiao H. Zhao X.W. Deng (2005) ArticleTitleOrgan-specific expression of Arabidopsis genome during development Plant Physiol. 138 80–91 Occurrence Handle10.1104/pp.104.054783 Occurrence Handle15888681
K. Marshall K. Nash G. Haussman I. Cassady D. Hume J. Jersey Particlede S. Hamilton (1997) ArticleTitleRecombinant human and mouse purple acid phosphatases: expression and characterization Arch. Biochem. Biophys. 345 230–236 Occurrence Handle10.1006/abbi.1997.0250 Occurrence Handle9308894
S.S. Miller J. Liu D.L. Allan C.J. Menzhuber M. Fedorova C.P. Vance (2001) ArticleTitleMolecular control of acid phosphatase secretion into the rhizosphere of proteoid from phosphorus-stressed white lupin Plant Physiol. 127 594–606 Occurrence Handle10.1104/pp.127.2.594 Occurrence Handle11598233
E. Moyano J.F. Martinez-Garcia C. Martin (1996) ArticleTitleApparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in Antirrhinum flowers Plant Cell 8 1519–1532 Occurrence Handle10.1105/tpc.8.9.1519 Occurrence Handle8837506
H. Nakazato T. Okamoto M. Nishikoori K. Washio N. Morita K. Haraguchi G.A. Thompson SuffixJr. H. Okuyama (1998) ArticleTitleThe glycosylphosphatidylinositol-anchored phosphatase from Spirodela oligorrhiza is a purple acid phosphatase Plant Physiol. 118 1015–1020 Occurrence Handle10.1104/pp.118.3.1015 Occurrence Handle9808746
J. Normanly B. Bartel (1999) ArticleTitleRedundancy as a way of life: IAA metabolism Curr. Opin. Plant Biol. 2 207–213 Occurrence Handle10.1016/S1369-5266(99)80037-5 Occurrence Handle10375566
F.B. Pickett D.R. Meeks-Wagner (1995) ArticleTitleSeeing Double: Appreciating Genetic Redundancy Plant Cell 7 1347–1356 Occurrence Handle10.1105/tpc.7.9.1347 Occurrence Handle8589620
R.H. Renegar F.W. Bazer R.M. Roberts (1982) ArticleTitlePlacental transport and distribution of uteroferrin in the fetal pig Biol. Reprod 27 1247–1260 Occurrence Handle7159665
J. Sambrook D.W. Russell (2001) Molecular Cloning EditionNumber3 Cold Spring Harbor Press Cold Spring Harbor, New York
G. Schenk Y. Ge L.E. Carrington C.J. Wynne I.R. Searle B.J. Carroll S. Hamilton J. Jersey Particlede (1999) ArticleTitleBinuclear metal centers in plant purple acid phosphatases: Fe–Mn in sweet potato and Fe–Zn in soybean Arch. Biochem. Biophys. 370 183–189 Occurrence Handle10.1006/abbi.1999.1407 Occurrence Handle10510276
G. Schenk M.L.J. Korsinczky D.A. Hume S. Hamilton J. Jersey Particlede (2000a) ArticleTitlePurple acid phosphatases from bacteria: similarities to mammalian and plant enzymes Gene 255 419–424 Occurrence Handle10.1016/S0378-1119(00)00305-X
G. Schenk Guddat L.w. Y. Ge L.E. Carrington D.A. Hume S. Hamilton J. Jersey Particlede (2000b) ArticleTitleIdentification of mammalian-like purple acid phosphatases in a wide range of plants Gene 250 117–125 Occurrence Handle10.1016/S0378-1119(00)00186-4
D.R. Smyth J.L. Bowman E.M. Meyerowitz (1990) ArticleTitleEarly flower development in Arabidopsis Plant cell 2 755–767 Occurrence Handle10.1105/tpc.2.8.755 Occurrence Handle2152125
N. Sträter T. Klabunde P. Tucker H. Witzel B. Krebs (1995) ArticleTitleCrystal structure of a purple acid phosphatase containing a dinuclear Fe(III)–Zn(II) active site Science 268 1489–1492 Occurrence Handle7770774
J.S. Taylor J. Raes (2004) ArticleTitleDuplication and divergence: the evolution of new genes and old ideas Annu. Rev. Genet. 38 615–643 Occurrence Handle10.1146/annurev.genet.38.072902.092831 Occurrence Handle15568988
F.A. Engelen Particlevan J.W. Molthoff A.J. Conner J.P. Nap A. Pereira W.J. Stiekema (1995) ArticleTitlepBINPLUS: an improved plant transformation vector based on pBIN19 Transgenic Res. 4 288–290 Occurrence Handle10.1007/BF01969123 Occurrence Handle7655517
J.B. Vincent B.A. Averill (1990) ArticleTitleAn enzyme with a double identity: purple acid phosphatase and tartrate-resistant acid phosphatase FASEB. J. 4 3009–3014 Occurrence Handle2394317
J.B. Vincent G.L. Oliver-Lilly B.A. Averill (1995) ArticleTitleProteins containing oxo-bridged dinuclear centers: a bioinorganic perspective Chem. Rev. 90 1447–1467 Occurrence Handle10.1021/cr00106a004
A. Vogel T. Borchers K. Marcus H.E. Meyer B. Krebs F. Spener (2002) ArticleTitleHeterologous expression and characterization of recombinant purple acid phosphatase from red kidney bean Arch. Biochem. Biophys. 401 164–172 Occurrence Handle10.1016/S0003-9861(02)00046-2 Occurrence Handle12054466
F. Wellmer J.L. Riechmann M. Alves-Ferreira E.M. Meyerowitz (2004) ArticleTitleGenome-wide analysis of spatial gene expression in Arabidopsis flowers Plant Cell 16 1314–1326 Occurrence Handle10.1105/tpc.021741 Occurrence Handle15100403
S.V. Wesley C.A. Helliwell N.A. Smith M.B. Wang D.T. Rouse Q. Liu P.S. Gooding S.P. Singh D. Abbott P.A. Stoutjesdijk (2001) ArticleTitleConstruct design for efficient, effective and high-throughput gene silencing in plants Plant J. 27 581–590 Occurrence Handle10.1046/j.1365-313X.2001.01105.x Occurrence Handle11576441
G.A. Wray M.W. Hahn E. Abouheif J.P. Balhoff M. Pizer M.V. Rockman L.A. Romano (2003) ArticleTitleThe evolution of transcriptional regulation in eukaryotes Mol. Biol. Evol. 20 1377–1419 Occurrence Handle10.1093/molbev/msg140 Occurrence Handle12777501
Y.L. Xiao M. Malik C.A. Whitelaw C.D. Town (2002) ArticleTitleCloning and sequencing of cDNAs for hypothetical genes from chromosome 2 of Arabidopsis Plant Physiol. 130 2118–2128 Occurrence Handle10.1104/pp.010207 Occurrence Handle12481096
P. Zimmermann B. Regierer J. Kossmann E. Frossard N. Amrhein M Bucher (2004) ArticleTitleDifferential expression of three purple acid phosphatases from potato Plant Biol. (Stuttg) 6 519–528 Occurrence Handle10.1055/s-2004-821091
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Zhu, H., Qian, W., Lu, X. et al. Expression Patterns of Purple Acid Phosphatase Genes in Arabidopsis Organs and Functional Analysis of AtPAP23 Predominantly Transcribed in Flower. Plant Mol Biol 59, 581–594 (2005). https://doi.org/10.1007/s11103-005-0183-0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11103-005-0183-0