Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Site Tools

  • AAAS
  • Subscribe
  • Feedback

Site Search

Search Advanced

Logo for

Genes & Dev. 14 (4): 435-451

Copyright © 2000 by Cold Spring Harbor Laboratory Press.

Vol. 14, No. 4, pp. 435-451, February 15, 2000

RESEARCH PAPER
Homeodomain and winged-helix transcription factors recruit activated Smads to distinct promoter elements via a common Smad interaction motif

Stéphane Germain,1,4 Michael Howell,1,2 Graeme M. Esslemont,2 and Caroline S. Hill1,2,3

1 Laboratory of Developmental Signalling, Imperial Cancer Research Fund, WC2A 3PX London, UK; 2 Ludwig Institute for Cancer Research, W1P 6BT London, UK

We have investigated the regulation of the activin-inducible distal element (DE) of the Xenopus goosecoid promoter. The results show that paired-like homeodomain transcription factors of the Mix family, Mixer and Milk, but not Mix.1, mediate activin/TGF-beta -induced transcription through the DE by interacting with the effector domain of Smad2, thereby recruiting active Smad2/Smad4 complexes to the Mixer/Milk-binding site. We identify a short motif in the carboxyl termini of Mixer and Milk, which is demonstrated to be both necessary and sufficient for interaction with the effector domain of Smad2 and is required for mediating activin/TGF-beta -induced transcription. This motif is not confined to these homeodomain proteins, but is also present in the Smad2-interacting winged-helix proteins Xenopus Fast-1, human Fast-1, and mouse Fast-2. We demonstrate directly that transcription factors of different DNA-binding specificity recruit activated Smads to distinct promoter elements via a common mechanism. These observations, together with the temporal and spatial expression patterns of Mixer and Milk, lead us to propose a model for mesoendoderm formation in Xenopus in which these homeodomain transcription factor/Smad complexes play a role in initiating and maintaining transcription of target genes in response to endogenous activin-like signals.

[Key Words: Activin; homeodomain; Smad; TGF-beta ; transcriptional regulation; Xenopus]

4 Present address: INSERM Unité 36, Collège de France, 3 rue d'Ulm, 75005 Paris, France.

3 Corresponding author.

GENES & DEVELOPMENT 14:435-451 © 2000 by Cold Spring Harbor Laboratory Press  ISSN 0890-9369/00 $5.00

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Transforming Growth Factor {beta}-Induced Smad1/5 Phosphorylation in Epithelial Cells Is Mediated by Novel Receptor Complexes and Is Essential for Anchorage-Independent Growth.
A. C. Daly, R. A. Randall, and C. S. Hill (2008)
Mol. Cell. Biol. 28, 6889-6902
   Abstract »    Full Text »    PDF »
The pro-domain of the zebrafish Nodal-related protein Cyclops regulates its signaling activities.
J. Tian, B. Andree, C. M. Jones, and K. Sampath (2008)
Development 135, 2649-2658
   Abstract »    Full Text »    PDF »
Mathematical modeling identifies Smad nucleocytoplasmic shuttling as a dynamic signal-interpreting system.
B. Schmierer, A. L. Tournier, P. A. Bates, and C. S. Hill (2008)
PNAS 105, 6608-6613
   Abstract »    Full Text »    PDF »
Chromatin-Bound p53 Anchors Activated Smads and the mSin3A Corepressor To Confer Transforming Growth Factor {beta}-Mediated Transcription Repression.
D. S. Wilkinson, W.-W. Tsai, M. A. Schumacher, and M. C. Barton (2008)
Mol. Cell. Biol. 28, 1988-1998
   Abstract »    Full Text »    PDF »
Transforming growth factor-{beta} signaling and ubiquitinators in cancer.
E. Glasgow and L. Mishra (2008)
Endocr. Relat. Cancer 15, 59-72
   Abstract »    Full Text »    PDF »
Erbin Inhibits Transforming Growth Factor {beta} Signaling through a Novel Smad-Interacting Domain.
F. Dai, C. Chang, X. Lin, P. Dai, L. Mei, and X.-H. Feng (2007)
Mol. Cell. Biol. 27, 6183-6194
   Abstract »    Full Text »    PDF »
Arkadia Activates Smad3/Smad4-Dependent Transcription by Triggering Signal-Induced SnoN Degradation.
L. Levy, M. Howell, D. Das, S. Harkin, V. Episkopou, and C. S. Hill (2007)
Mol. Cell. Biol. 27, 6068-6083
   Abstract »    Full Text »    PDF »
Nodal signaling: developmental roles and regulation.
M. M. Shen (2007)
Development 134, 1023-1034
   Abstract »    Full Text »    PDF »
Sequence comparison by sequence harmony identifies subtype-specific functional sites.
W. Pirovano, K. A. Feenstra, and J. Heringa (2006)
Nucleic Acids Res. 34, 6540-6548
   Abstract »    Full Text »    PDF »
The Carboxyl-terminal Nucleoplasmic Region of MAN1 Exhibits a DNA Binding Winged Helix Domain.
S. Caputo, J. Couprie, I. Duband-Goulet, E. Konde, F. Lin, S. Braud, M. Gondry, B. Gilquin, H. J. Worman, and S. Zinn-Justin (2006)
J. Biol. Chem. 281, 18208-18215
   Abstract »    Full Text »    PDF »
Zebrafish endoderm formation is regulated by combinatorial Nodal, FGF and BMP signalling.
M. Poulain, M. Furthauer, B. Thisse, C. Thisse, and T. Lepage (2006)
Development 133, 2189-2200
   Abstract »    Full Text »    PDF »
Requirement for the SnoN Oncoprotein in Transforming Growth Factor {beta}-Induced Oncogenic Transformation of Fibroblast Cells.
Q. Zhu, S. Pearson-White, and K. Luo (2005)
Mol. Cell. Biol. 25, 10731-10744
   Abstract »    Full Text »    PDF »
Smad transcription factors.
J. Massague, J. Seoane, and D. Wotton (2005)
Genes & Dev. 19, 2783-2810
   Abstract »    Full Text »    PDF »
Kinetic Analysis of Smad Nucleocytoplasmic Shuttling Reveals a Mechanism for Transforming Growth Factor {beta}-Dependent Nuclear Accumulation of Smads.
B. Schmierer and C. S. Hill (2005)
Mol. Cell. Biol. 25, 9845-9858
   Abstract »    Full Text »    PDF »
Maternal Xenopus Zic2 negatively regulates Nodal-related gene expression during anteroposterior patterning.
D. W. Houston and C. Wylie (2005)
Development 132, 4845-4855
   Abstract »    Full Text »    PDF »
Bifurcated converging pathways for high Ca2+- and TGF{beta}-induced inhibition of growth of normal human keratinocytes.
M. Sakaguchi, H. Sonegawa, T. Nukui, Y. Sakaguchi, M. Miyazaki, M. Namba, and N.-h. Huh (2005)
PNAS 102, 13921-13926
   Abstract »    Full Text »    PDF »
Smad4 Dependency Defines Two Classes of Transforming Growth Factor {beta} (TGF-{beta}) Target Genes and Distinguishes TGF-{beta}-Induced Epithelial-Mesenchymal Transition from Its Antiproliferative and Migratory Responses.
L. Levy and C. S. Hill (2005)
Mol. Cell. Biol. 25, 8108-8125
   Abstract »    Full Text »    PDF »
Positive and Negative Regulation of the Transforming Growth Factor {beta}/Activin Target Gene goosecoid by the TFII-I Family of Transcription Factors.
M. Ku, S. Y. Sokol, J. Wu, M. I. Tussie-Luna, A. L. Roy, and A. Hata (2005)
Mol. Cell. Biol. 25, 7144-7157
   Abstract »    Full Text »    PDF »
C-terminal mutants of C. elegans Smads reveal tissue-specific requirements for protein activation by TGF-{beta} signaling.
J. Wang, W. A. Mohler, and C. Savage-Dunn (2005)
Development 132, 3505-3513
   Abstract »    Full Text »    PDF »
Bone Morphogenetic Proteins in Vascular Calcification.
K. A. Hruska, S. Mathew, and G. Saab (2005)
Circ. Res. 97, 105-114
   Abstract »    Full Text »    PDF »
TGF-{beta} signaling potentiates differentiation of embryonic stem cells to Pdx-1 expressing endodermal cells.
N. Shiraki, C.-J. Lai, Y. Hishikari, and S. Kume (2005)
Genes Cells 10, 503-516
   Abstract »    Full Text »    PDF »
Bone Morphogenic Protein (Smad)-Mediated Repression of Proopiomelanocortin Transcription by Interference with Pitx/Tpit Activity.
M. Nudi, J.-F. Ouimette, and J. Drouin (2005)
Mol. Endocrinol. 19, 1329-1342
   Abstract »    Full Text »    PDF »
Gene Regulatory Networks Special Feature: Xenopus as a model system to study transcriptional regulatory networks.
T. Koide, T. Hayata, and K. W. Y. Cho (2005)
PNAS 102, 4943-4948
   Abstract »    Full Text »    PDF »
Activin Responsiveness of the Murine Gonadotropin-Releasing Hormone Receptor Gene Is Mediated by a Composite Enhancer Containing Spatially Distinct Regulatory Elements.
B. D. Cherrington, T. A. Farmerie, C. A. Lents, J. D. Cantlon, M. S. Roberson, and C. M. Clay (2005)
Mol. Endocrinol. 19, 898-912
   Abstract »    Full Text »    PDF »
SnoN Is a Cell Type-specific Mediator of Transforming Growth Factor-{beta} Responses.
K. P. Sarker, S. M. Wilson, and S. Bonni (2005)
J. Biol. Chem. 280, 13037-13046
   Abstract »    Full Text »    PDF »
GATA4, 5 and 6 mediate TGF{beta} maintenance of endodermal gene expression in Xenopus embryos.
B. A. Afouda, A. Ciau-Uitz, and R. Patient (2005)
Development 132, 763-774
   Abstract »    Full Text »    PDF »
A Direct Intersection between p53 and Transforming Growth Factor {beta} Pathways Targets Chromatin Modification and Transcription Repression of the {alpha}-Fetoprotein Gene.
D. S. Wilkinson, S. K. Ogden, S. A. Stratton, J. L. Piechan, T. T. Nguyen, G. A. Smulian, and M. C. Barton (2005)
Mol. Cell. Biol. 25, 1200-1212
   Abstract »    Full Text »    PDF »
Negative regulation of Smad2 by PIASy is required for proper Xenopus mesoderm formation.
M. Daniels, K. Shimizu, A. M. Zorn, and S.-i. Ohnuma (2004)
Development 131, 5613-5626
   Abstract »    Full Text »    PDF »
New roles for FoxH1 in patterning the early embryo.
M. Kofron, H. Puck, H. Standley, C. Wylie, R. Old, M. Whitman, and J. Heasman (2004)
Development 131, 5065-5078
   Abstract »    Full Text »    PDF »
Disorder in a Target for the Smad2 Mad Homology 2 Domain and Its Implications for Binding and Specificity.
P. A. Chong, B. Ozdamar, J. L. Wrana, and J. D. Forman-Kay (2004)
J. Biol. Chem. 279, 40707-40714
   Abstract »    Full Text »    PDF »
Analysis of Smad nucleocytoplasmic shuttling in living cells.
F. J. Nicolas, K. De Bosscher, B. Schmierer, and C. S. Hill (2004)
J. Cell Sci. 117, 4113-4125
   Abstract »    Full Text »    PDF »
Opposite Smad and Chicken Ovalbumin Upstream Promoter Transcription Factor Inputs in the Regulation of the Collagen VII Gene Promoter by Transforming Growth Factor-{beta}.
M. J. Calonge, J. Seoane, and J. Massague (2004)
J. Biol. Chem. 279, 23759-23765
   Abstract »    Full Text »    PDF »
The role of Mixer in patterning the early Xenopus embryo.
M. Kofron, C. Wylie, and J. Heasman (2004)
Development 131, 2431-2441
   Abstract »    Full Text »    PDF »
Activin Regulation of the Follicle-Stimulating Hormone {beta}-Subunit Gene Involves Smads and the TALE Homeodomain Proteins Pbx1 and Prep1.
J. S. Bailey, N. Rave-Harel, S. M. McGillivray, D. Coss, and P. L. Mellon (2004)
Mol. Endocrinol. 18, 1158-1170
   Abstract »    Full Text »    PDF »
Recognition of Phosphorylated-Smad2-Containing Complexes by a Novel Smad Interaction Motif.
R. A. Randall, M. Howell, C. S. Page, A. Daly, P. A. Bates, and C. S. Hill (2004)
Mol. Cell. Biol. 24, 1106-1121
   Abstract »    Full Text »    PDF »
Mixer/Bon and FoxH1/Sur have overlapping and divergent roles in Nodal signaling and mesendoderm induction.
P. S. Kunwar, S. Zimmerman, J. T. Bennett, Y. Chen, M. Whitman, and A. F. Schier (2003)
Development 130, 5589-5599
   Abstract »    Full Text »    PDF »
The Mix family homeodomain gene bonnie and clyde functions with other components of the Nodal signaling pathway to regulate neural patterning in zebrafish.
L. A. Trinh, D. Meyer, and D. Y. R. Stainier (2003)
Development 130, 4989-4998
   Abstract »    Full Text »    PDF »
Regulation of apoptosis in theXenopus embryo by Bix3.
M. Trindade, N. Messenger, C. Papin, D. Grimmer, L. Fairclough, M. Tada, and J. C. Smith (2003)
Development 130, 4611-4622
   Abstract »    Full Text »    PDF »
Interplay between the tumor suppressor p53 and TGF{beta} signaling shapes embryonic body axes in Xenopus.
K. Takebayashi-Suzuki, J. Funami, D. Tokumori, A. Saito, T. Watabe, K. Miyazono, A. Kanda, and A. Suzuki (2003)
Development 130, 3929-3939
   Abstract »    Full Text »    PDF »
The Transforming Activity of Ski and SnoN Is Dependent on Their Ability to Repress the Activity of Smad Proteins.
J. He, S. B. Tegen, A. R. Krawitz, G. S. Martin, and K. Luo (2003)
J. Biol. Chem. 278, 30540-30547
   Abstract »    Full Text »    PDF »
Joint regulation of the MAP1B promoter by HNF3{beta}/Foxa2 and Engrailed is the result of a highly conserved mechanism for direct interaction of homeoproteins and Fox transcription factors.
I. Foucher, M. L. Montesinos, M. Volovitch, A. Prochiantz, and A. Trembleau (2003)
Development 130, 1867-1876
   Abstract »    Full Text »    PDF »
The Foxh1-dependent autoregulatory enhancer controls the level of Nodal signals in the mouse embryo.
D. P. Norris, J. Brennan, E. K. Bikoff, and E. J. Robertson (2003)
Development 129, 3455-3468
   Abstract »    Full Text »    PDF »
A novel Xenopus Smad-interacting forkhead transcription factor (XFast-3) cooperates with XFast-1 in regulating gastrulation movements.
M. Howell, G. J. Inman, and C. S. Hill (2003)
Development 129, 2823-2834
   Abstract »    Full Text »    PDF »
A study of mesoderm patterning through the analysis of the regulation of Xmyf-5 expression.
M. Polli and E. Amaya (2003)
Development 129, 2917-2927
   Abstract »    Full Text »    PDF »
Two Short Segments of Smad3 Are Important for Specific Interaction of Smad3 with c-Ski and SnoN.
M. Mizuide, T. Hara, T. Furuya, M. Takeda, K. Kusanagi, Y. Inada, M. Mori, T. Imamura, K. Miyazawa, and K. Miyazono (2003)
J. Biol. Chem. 278, 531-536
   Abstract »    Full Text »    PDF »
Stoichiometry of Active Smad-Transcription Factor Complexes on DNA.
G. J. Inman and C. S. Hill (2002)
J. Biol. Chem. 277, 51008-51016
   Abstract »    Full Text »    PDF »
Transforming Growth Factor-beta Inhibits Pulmonary Surfactant Protein B Gene Transcription through SMAD3 Interactions with NKX2.1 and HNF-3 Transcription Factors.
C. Li, N.-L. Zhu, R. C. Tan, P. L. Ballard, R. Derynck, and P. Minoo (2002)
J. Biol. Chem. 277, 38399-38408
   Abstract »    Full Text »    PDF »
Mixl1 is required for axial mesendoderm morphogenesis and patterning in the murine embryo.
A. H. Hart, L. Hartley, K. Sourris, E. S. Stadler, R. Li, E. G. Stanley, P. P. L. Tam, A. G. Elefanty, and L. Robb (2002)
Development 129, 3597-3608
   Abstract »    Full Text »    PDF »
Bone morphogenetic protein-4-induced activation of Xretpos is mediated by Smads and Olf-1/EBF associated zinc finger (OAZ).
S. Shim, N. Bae, and J.-K. Han (2002)
Nucleic Acids Res. 30, 3107-3117
   Abstract »    Full Text »    PDF »
SB-431542 Is a Potent and Specific Inhibitor of Transforming Growth Factor-beta Superfamily Type I Activin Receptor-Like Kinase (ALK) Receptors ALK4, ALK5, and ALK7.
G. J. Inman, F. J. Nicolas, J. F. Callahan, J. D. Harling, L. M. Gaster, A. D. Reith, N. J. Laping, and C. S. Hill (2002)
Mol. Pharmacol. 62, 65-74
   Abstract »    Full Text »    PDF »
Transforming growth factor {beta} signal transduction.
S. Dennler, M.-J. Goumans, and P. ten Dijke (2002)
J. Leukoc. Biol. 71, 731-740
   Abstract »    Full Text »    PDF »
A glimpse into the molecular entrails of endoderm formation.
D. Y.R. Stainier (2002)
Genes & Dev. 16, 893-907
   Full Text »    PDF »
The role of a Williams-Beuren syndrome-associated helix-loop-helix domain-containing transcription factor in activin/nodal signaling.
C. Ring, S. Ogata, L. Meek, J. Song, T. Ohta, K. Miyazono, and K. W.Y. Cho (2002)
Genes & Dev. 16, 820-835
   Abstract »    Full Text »    PDF »
Transforming Growth Factor-beta Induction of Smooth Muscle Cell Phenotpye Requires Transcriptional and Post-transcriptional Control of Serum Response Factor.
K. K. Hirschi, L. Lai, N. S. Belaguli, D. A. Dean, R. J. Schwartz, and W. E. Zimmer (2002)
J. Biol. Chem. 277, 6287-6295
   Abstract »    Full Text »    PDF »
Mezzo, a paired-like homeobox protein is an immediate target of Nodal signalling and regulates endoderm specification in zebrafish.
M. Poulain and T. Lepage (2002)
Development 129, 4901-4914
   Abstract »    Full Text »    PDF »
A changing morphogen gradient is interpreted by continuous transduction flow.
P.-Y. Bourillot, N. Garrett, and J. B. Gurdon (2002)
Development 129, 2167-2180
   Abstract »    Full Text »    PDF »
Meeting Report: Signaling Schemes for TGF-{beta}.
A. B. Roberts and R. Derynck (2001)
Science Signaling 2001 , pe43
   Abstract »    Full Text »    PDF »
Molecular effects of novel mutations in Hesx1/HESX1 associated with human pituitary disorders.
J. M. Brickman, M. Clements, R. Tyrell, D. McNay, K. Woods, J. Warner, A. Stewart, R. S. P. Beddington, and M. Dattani (2001)
Development 128, 5189-5199
   Abstract »    Full Text »    PDF »
Loss of Smad4 Function in Pancreatic Tumors. C-TERMINAL TRUNCATION LEADS TO DECREASED STABILITY.
D. Maurice, C. E. Pierreux, M. Howell, R. E. Wilentz, M. J. Owen, and C. S. Hill (2001)
J. Biol. Chem. 276, 43175-43181
   Abstract »    Full Text »    PDF »
Smad3 recruits the anaphase-promoting complex for ubiquitination and degradation of SnoN.
S. L. Stroschein, S. Bonni, J. L. Wrana, and K. Luo (2001)
Genes & Dev. 15, 2822-2836
   Abstract »    Full Text »    PDF »
Swift Is a Novel BRCT Domain Coactivator of Smad2 in Transforming Growth Factor {beta} Signaling.
K. Shimizu, P.-Y. Bourillot, S. J. Nielsen, A. M. Zorn, and J. B. Gurdon (2001)
Mol. Cell. Biol. 21, 3901-3912
   Abstract »    Full Text »
FoxH1 (Fast) functions to specify the anterior primitive streak in the mouse.
P. A. Hoodless, M. Pye, C. Chazaud, E. Labbé, L. Attisano, J. Rossant, and J. L. Wrana (2001)
Genes & Dev. 15, 1257-1271
   Abstract »    Full Text »
Smad proteins function as co-modulators for MEF2 transcriptional regulatory proteins.
Z. A. Quinn, C.-C. Yang, J. L. Wrana, and J. C. McDermott (2001)
Nucleic Acids Res. 29, 732-742
   Abstract »    Full Text »    PDF »
Multiple roles for Gata5 in zebrafish endoderm formation.
J. Reiter, Y Kikuchi, and D. Stainier (2001)
Development 128, 125-135
   Abstract »    PDF »
Maternal VegT is the initiator of a molecular network specifying endoderm in Xenopus laevis.
J. Xanthos, M Kofron, C Wylie, and J Heasman (2001)
Development 128, 167-180
   Abstract »    PDF »
Arabidopsis Transcription Factors: Genome-Wide Comparative Analysis Among Eukaryotes.
J. L. Riechmann, J. Heard, G. Martin, L. Reuber, C. -Z., Jiang, J. Keddie, L. Adam, O. Pineda, O. J. Ratcliffe, et al. (2000)
Science 290, 2105-2110
   Abstract »    Full Text »
Transforming Growth Factor beta -Independent Shuttling of Smad4 between the Cytoplasm and Nucleus.
C. E. Pierreux, F. J. Nicolás, and C. S. Hill (2000)
Mol. Cell. Biol. 20, 9041-9054
   Abstract »    Full Text »
Raf induces TGFbeta production while blocking its apoptotic but not invasive responses: a mechanism leading to increased malignancy in epithelial cells.
K. Lehmann, E. Janda, C. E. Pierreux, M. Rytömaa, A. Schulze, M. McMahon, C. S. Hill, H. Beug, and J. Downward (2000)
Genes & Dev. 14, 2610-2622
   Abstract »    Full Text »
Role of Transforming Growth Factor-{beta} Signaling in Cancer.
M. P. de Caestecker, E. Piek, and A. B. Roberts (2000)
J Natl Cancer Inst 92, 1388-1402
   Abstract »    Full Text »    PDF »
BF-1 Interferes with Transforming Growth Factor beta Signaling by Associating with Smad Partners.
C. Dou, J. Lee, B. Liu, F. Liu, J. Massague, S. Xuan, and E. Lai (2000)
Mol. Cell. Biol. 20, 6201-6211
   Abstract »    Full Text »
Controlling TGF-beta signaling.
J. Massagué and Y.-G. Chen (2000)
Genes & Dev. 14, 627-644
   Full Text »
Formation of the definitive endoderm in mouse is a Smad2-dependent process.
K. Tremblay, P. Hoodless, E. Bikoff, and E. Robertson (2000)
Development 127, 3079-3090
   Abstract »    PDF »
Inhibition of the Transforming Growth Factor beta 1 Signaling Pathway by the AML1/ETO Leukemia-associated Fusion Protein.
A. Jakubowiak, C. Pouponnot, F. Berguido, R. Frank, S. Mao, J. Massague, and S. D. Nimer (2000)
J. Biol. Chem. 275, 40282-40287
   Abstract »    Full Text »    PDF »
Distinct Oligomeric States of SMAD Proteins in the Transforming Growth Factor-beta Pathway.
L. Jayaraman and J. Massague (2000)
J. Biol. Chem. 275, 40710-40717
   Abstract »    Full Text »    PDF »
Autoregulation of Xvent-2B; Direct Interaction and Functional Cooperation of Xvent-2 and Smad1.
K. A. Henningfeld, H. Friedle, S. Rastegar, and W. Knochel (2002)
J. Biol. Chem. 277, 2097-2103
   Abstract »    Full Text »    PDF »
Transforming growth factor beta -inducible independent binding of SMAD to the Smad7 promoter.
N. G. Denissova, C. Pouponnot, J. Long, D. He, and F. Liu (2000)
PNAS 97, 6397-6402
   Abstract »    Full Text »    PDF »

ADVERTISEMENT
Click Me!

ADVERTISEMENT
Click Me!

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (pre-2008: Science's STKE. ISSN 1525-8882)