Detection of APC mutations by a yeast-based protein truncation test (YPTT)

Takao Suzuki, Chikashi Ishioka, Satoshi Kato, Yasushi Mitachi, Hideki Shimodaira, Masato Sakayori, Akira Shimada, Mitsuo Asamura, Ryunosuke Kanamaru

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

APC gene mutations play a role in the initiation step of colorectal carcinogenesis in both familial adenomatous polyposis (FAP) and non-FAP patients. Almost all of the APC mutations are nonsense or frameshift mutations, which truncate the APC protein and are thought to inactivate normal APC function. We show a novel method for detecting nonsense and frameshift APC gene mutations by using Saccharomyces cerevisiae. Polymerase chain reaction (PCR)-amplified APC fragments are cloned directly into yeast expression vectors in vivo, and the yeast expresses a hemagglutinin epitope (HA)-tagged APC peptide. Widen an APC fragment contains a nonsense or frameshift mutation, HA-tagged truncating APC peptide can be detected by Western blotting using an anti-HA antibody. We identified both germ-line and somatic APC mutations in patients with FAP and non-FAP colorectal tumors, respectively. This method, called the yeast-based protein truncation test (YPTT), is simple and fairly cheap, and it can be applied to any genes that are inactivated by protein truncating mutations.

Original languageEnglish
Pages (from-to)290-297
Number of pages8
JournalGenes Chromosomes and Cancer
Volume21
Issue number4
DOIs
Publication statusPublished - 1998
Externally publishedYes

Fingerprint

Fungal Proteins
Hemagglutinins
Mutation
APC Genes
Epitopes
Frameshift Mutation
Adenomatous Polyposis Coli
Nonsense Codon
Yeasts
Peptides
Germ Cells
Saccharomyces cerevisiae
Colorectal Neoplasms
Carcinogenesis
Proteins
Western Blotting
Polymerase Chain Reaction
Antibodies

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Suzuki, T., Ishioka, C., Kato, S., Mitachi, Y., Shimodaira, H., Sakayori, M., ... Kanamaru, R. (1998). Detection of APC mutations by a yeast-based protein truncation test (YPTT). Genes Chromosomes and Cancer, 21(4), 290-297. https://doi.org/10.1002/(SICI)1098-2264(199804)21:4<290::AID-GCC2>3.0.CO;2-U

Detection of APC mutations by a yeast-based protein truncation test (YPTT). / Suzuki, Takao; Ishioka, Chikashi; Kato, Satoshi; Mitachi, Yasushi; Shimodaira, Hideki; Sakayori, Masato; Shimada, Akira; Asamura, Mitsuo; Kanamaru, Ryunosuke.

In: Genes Chromosomes and Cancer, Vol. 21, No. 4, 1998, p. 290-297.

Research output: Contribution to journalArticle

Suzuki, T, Ishioka, C, Kato, S, Mitachi, Y, Shimodaira, H, Sakayori, M, Shimada, A, Asamura, M & Kanamaru, R 1998, 'Detection of APC mutations by a yeast-based protein truncation test (YPTT)', Genes Chromosomes and Cancer, vol. 21, no. 4, pp. 290-297. https://doi.org/10.1002/(SICI)1098-2264(199804)21:4<290::AID-GCC2>3.0.CO;2-U
Suzuki, Takao ; Ishioka, Chikashi ; Kato, Satoshi ; Mitachi, Yasushi ; Shimodaira, Hideki ; Sakayori, Masato ; Shimada, Akira ; Asamura, Mitsuo ; Kanamaru, Ryunosuke. / Detection of APC mutations by a yeast-based protein truncation test (YPTT). In: Genes Chromosomes and Cancer. 1998 ; Vol. 21, No. 4. pp. 290-297.
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