Familial juvenile polyposis is an autosomal dominant disease characterized by a predisposition to hamartomatous polyps and gastrointestinal cancer. Here it is shown that a subset of juvenile polyposis families carry germ line mutations in the geneSMAD4 (also known as DPC4), located on chromosome 18q21.1, that encodes a critical cytoplasmic mediator in the transforming growth factor–β signaling pathway. The mutant SMAD4 proteins are predicted to be truncated at the carboxyl-terminus and lack sequences required for normal function. These results confirm an important role for SMAD4 in the development of gastrointestinal tumors.

Get full access to this article

View all available purchase options and get full access to this article.


Järvinen H. J., Franssila K. O., Gut25, 792 (1984); J. R. Jass, in Familial Adenomatous Polyposis and Other Polyposis Syndromes, R. K. S. Phillips, A. D. Spigelman, J. P. S. Thomson, Eds. (Edward Arnold, London, 1994), pp. 203–214; J. R. Howe, F. A. Mitros, R. W. Summers, in preparation. Estimates of gastrointestinal cancer vary widely because there have been few studies in large families with long-term follow-up.
E. D. Lynch, et al., Am. J. Hum. Genet. 61, 1254 (1997); Olschwang S., et al., Nature Genet. 18, 12 (1998).
Liaw D., et al., Nature Genet.16, 64 (1997); D. J. Marsh et al.,ibid., p. 333.
Marsh D. J., et al., Cancer Res.57, 5017 (1997); G. J. Riggins, S. R. Hamilton, K. W. Kinzler, B. Vogelstein, “Normal PTEN Gene in Juvenile Polyposis,” J. Neg. Obs. Gen. Oncol. [online]1, 1 (1997). Available at
Eng C., Ji H., Am. J. Hum. Genet.62, 1020 (1998).
J. R. Howe et al., ibid., p. 1129.
Eppert K., et al., Cell86, 543 (1996).
T. J. Stemper, T. H. Kent, R. W. Summers,Ann. Int. Med. 83, 639 (1975).
Cho K. R., et al., Genomics19, 525 (1994).
Primers were designed for exons 1 to 29 of DCCwith the Primer3 server ( and the published intron-exon boundaries (14). Primers for amplification of the SMAD4 gene have been described (27). PCR was performed in a 10-μl volume with 25 ng of DNA, 200 μM each of the deoxynucleotide triphosphates dGTP, dATP, dTTP, and dCTP, 1 μl of 10× buffer [100 mM tris-HCl (pH 8.3), 500 mM KCl, 15 mM MgCl2, 0.01% w/v gelatin], 2 pmol of each primer, and 0.25 units of Taq DNA polymerase. PCR was performed for 1 min at 94°C, 1 min at 55°C (or optimal annealing temperature), and 1 min at 72°C for 30 cycles. After amplification, 5 μl of stop solution (95% formamide, 10 mM NaOH, 0.05% bromophenol blue, 0.05% xylene cyanol) was added, and samples were heated to 95°C for 3 min, then loaded onto 6% nondenaturing polyacrylamide gels (with and without 10% glycerol). DNA was detected by silver staining. Informed consent for DNA studies was obtained from family members with the approval of the Institutional Review Board at the University of Iowa.
PCR products were subjected to electrophoresis through 2% agarose gels and stained with ethidium bromide to confirm the presence of a single band of the expected size. The products were isolated with the Qiaquick PCR purification kit (Qiagen, Santa Clarita, CA) and then sequenced with the ABI Prism Dye Terminator Cycle Sequencing kit (PE Applied Biosystems, Foster City, CA). Cycle sequencing included 1 cycle at 98°C for 5 min, followed by 30 cycles at 94°C for 10 s, 50°C for 5 s, and 60°C for 4 min. Individual PCR primers were used for sequencing the sense and antisense strands for each exon. Reactions were analyzed with an ABI model 373XL stretch fluorescent automated sequencer.
Exon 9 was PCR amplified and the gel products purified and ligated into the p-GEM-T Easy plasmid vector (Promega). JM109 cells were transformed with the vector and plated onto LB-ampicillin plates containing 0.5 mM isopropylthio-β- d -galactoside and 5-bromo-4-chloro-3-indolyl-β- d -galactoside (80 μg/ml). Recombinant clones were grown overnight at 37°C in LB-ampicillin (100 μg/ml) medium. Cells were harvested and lysed, and cycle sequencing was performed with DPC4S9 and AS9 primers.
Hahn S. A., et al., Science271, 350 (1996).
Schutte M., et al., Cancer Res.56, 2527 (1996).
Thiagalingam S., et al., Nature Genet.13, 343 (1996).
Wrana J. L., Attisano L., Trends Genet.12, 493 (1996).
Lagna G., et al., Nature383, 832 (1996).
Wrana J., Pawson T., ibid388, 28 (1997).
Grau A. M., et al., Cancer Res.57, 3929 (1997).
Shi Y., et al., Nature388, 87 (1997).
Takagi Y., et al., Gastroenterology111, 1369 (1996).
Kim S. K., et al., Cancer Res.56, 2519 (1996).
Takaku K., et al., Cell92, 645 (1998).
L. A. Aaltonen, R. S. Houlston, S. Bevan, I. P. M. Tomlinson, unpublished data.
A. Hemminki et al., Nature 391, 184 (1998).
Li D.-M., Sun H., Cancer Res.57, 2124 (1997).
Moskaluk C. A., et al., Diagn. Mol. Pathol.6, 85 (1997).
Howe J. R., Klimstra D. S., Cordon-Cardo C., Histol. Histopathol.12, 595 (1997).
We thank A. Hemminki, P. Kristo, A. Loukola, E. Avizienyte, R. Salovaara, K. Saastamoinen, S. Lindh, T. Lehtinen, and T. Kosonen for assistance. Supported in part by the Owen H. Wangensteen Faculty Research Fellowship of the American College of Surgeons, the Carver Trust Medical Research Initiative Grant, the Clinical Cancer Center at the University of Iowa, the Academy of Finland, the Helsinki University Central Hospital, the Finnish Cancer Society, the Research and Science Foundation of Farmos, the European Commission, the Sigrid Juselius Foundation, and the Coeliac Disease Society.


eLetters is an online forum for ongoing peer review. Submission of eLetters are open to all. eLetters are not edited, proofread, or indexed. Please read our Terms of Service before submitting your own eLetter.

Log In to Submit a Response

No eLetters have been published for this article yet.

Information & Authors


Published In

Volume 280 | Issue 5366
15 May 1998

Submission history

Received: 17 March 1998
Accepted: 7 April 1998
Published in print: 15 May 1998


Request permissions for this article.



J. R. Howe and J. C. Ringold, Department of Surgery, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
S. Roth and L. A. Aaltonen, Department of Medical Genetics, Haartman Institute, FIN-00014, University of Helsinki, Finland.
R. W. Summers, Department of Medicine, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
H. J. Järvinen, Second Department of Surgery, Helsinki University Central Hospital, Haartmaninkatu 4, 00290 Helsinki, Finland.
P. Sistonen, Finnish Red Cross Blood Transfusion Service, Kivihaantie 7, 00130 Helsinki, Finland.
I. P. M. Tomlinson, Molecular and Population Genetics Laboratory, Imperial Cancer Research Fund, 44, Lincoln's Inn Fields, London, WC2A 3PX, UK.
R. S. Houlston and S. Bevan, Section of Cancer Genetics, Institute of Cancer Research, Sutton, Surrey SM2 5NG, UK.
F. A. Mitros, Department of Pathology, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
E. M. Stone, Department of Ophthalmology, University of Iowa College of Medicine, Iowa City, IA 52242, USA.


To whom correspondences should be addressed. E-mail: [email protected]

Metrics & Citations


Article Usage


Export citation

Select the format you want to export the citation of this publication.

Cited by


    View Options

    Check Access

    Log in to view the full text


    AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.

    Log in via OpenAthens.
    Log in via Shibboleth.
    More options

    Purchase digital access to this article

    Download and print this article for your personal scholarly, research, and educational use.

    Purchase this issue in print

    Buy a single issue of Science for just $15 USD.

    View options

    PDF format

    Download this article as a PDF file

    Download PDF

    Full Text








    Share article link

    Share on social media