A zebrafish bmyb mutation causes genome instability and increased cancer susceptibility.

A major goal of cancer research has been to identify genes that contribute to cancer formation. The similar pathology between zebrafish and human tumors, as well as the past success of large-scale genetic screens in uncovering human disease genes, makes zebrafish an ideal system in which to find such new genes. Here, we show that a zebrafish forward genetic screen uncovered multiple cell proliferation mutants including one mutant, crash&burn (crb), that represents a loss-of-function mutation in bmyb, a transcriptional regulator and member of a putative proto-oncogene family. crb mutant embryos have defects in mitotic progression and spindle formation, and exhibit genome instability. Regulation of cyclin B levels by bmyb appears to be the mechanism of mitotic accumulation in crb. Carcinogenesis studies reveal increased cancer susceptibility in adult crb heterozygotes. Gene-expression signatures associated with loss of bmyb in zebrafish are also correlated with conserved signatures in human tumor samples, and down-regulation of the B-myb signature genes is associated with retention of p53 function. Our findings show that zebrafish screens can uncover cancer pathways, and demonstrate that loss of function of bmyb is associated with cancer.

[1]  James F Amatruda,et al.  Zebrafish as a cancer model system. , 2002, Cancer cell.

[2]  R. Nairn,et al.  A CDKN2-like polymorphism in Xiphophorus LG V is associated with UV-B-induced melanoma formation in platyfish-swordtail hybrids. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J. Lipsick,et al.  Mutation of the Drosophila homologue of the Myb protooncogene causes genomic instability , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  D. Johnson,et al.  Increased E2F1 activity induces skin tumors in mice heterozygous and nullizygous for p53. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[5]  T. Poggio,et al.  Multiclass cancer diagnosis using tumor gene expression signatures , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[6]  L. Zon,et al.  tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. Watson,et al.  B‐Myb protein in cellular proliferation, transcription control, and cancer: Latest developments , 1999, Journal of cellular physiology.

[8]  E. Dı́az-Rodrı́guez,et al.  Complete loss of the tumor suppressor MAD2 causes premature cyclin B degradation and mitotic failure in human somatic cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[9]  T. Kornberg,et al.  Isolation of the proto-oncogene c-myb from D. melanogaster , 1985, Cell.

[10]  J. Postlethwait,et al.  Production of haploid and diploid androgenetic zebrafish (including methodology for delayed in vitro fertilization). , 1999, Methods in Cell Biology.

[11]  A. Krämer,et al.  Centrosome replication, genomic instability and cancer , 2002, Leukemia.

[12]  G. Ramsay,et al.  Mutations in Drosophila myb lead to centrosome amplification and genomic instability. , 2002, Development.

[13]  S. Ishii,et al.  Myb controls G2/M progression by inducing cyclin B expression in the Drosophila eye imaginal disc , 2002, The EMBO journal.

[14]  S. Ekker,et al.  Effective targeted gene ‘knockdown’ in zebrafish , 2000, Nature Genetics.

[15]  E. Lam,et al.  Characterization and cell cycle-regulated expression of mouse B-myb. , 1992, Oncogene.

[16]  T. Jacks,et al.  Loss of E2F-1 reduces tumorigenesis and extends the lifespan of Rb1(+/−) mice , 1998, Nature Genetics.

[17]  J. Doonan,et al.  G2/M-Phase–Specific Transcription during the Plant Cell Cycle Is Mediated by c-Myb–Like Transcription Factors , 2001, The Plant Cell Online.

[18]  J. Nevins,et al.  E2Fs link the control of G1/S and G2/M transcription , 2004, The EMBO journal.

[19]  Nancy Hopkins,et al.  Many Ribosomal Protein Genes Are Cancer Genes in Zebrafish , 2004, PLoS biology.

[20]  M. Daly,et al.  PGC-1α-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes , 2003, Nature Genetics.

[21]  David M Langenau,et al.  Myc-Induced T Cell Leukemia in Transgenic Zebrafish , 2003, Science.

[22]  J. Lipsick.  synMuv verite--Myb comes into focus. , 2004, Genes & development.

[23]  L. Zon,et al.  BRAF Mutations Are Sufficient to Promote Nevi Formation and Cooperate with p53 in the Genesis of Melanoma , 2005, Current Biology.

[24]  A. Schier,et al.  Efficient recovery of ENU-induced mutations from the zebrafish germline. , 1994, Genetics.

[25]  R. Watson,et al.  Cell cycle regulation by the B-Myb transcription factor , 2003, Cellular and Molecular Life Sciences CMLS.

[26]  J. Adolfsson,et al.  Detecting homozygous deletions in the CDKN2A(p16(INK4a))/ARF(p14(ARF)) gene in urinary bladder cancer using real-time quantitative PCR. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[27]  J. Hendricks,et al.  Neoplasia in Zebrafish (Danio rerio) Treated with N-methyl-N'nitro-N-nitrosoguanidine by Three Exposure Routes at ifferent Developmental Stages , 2000, Toxicologic pathology.

[28]  Masaki Ito,et al.  Factors controlling cyclin B expression , 2000, Plant Molecular Biology.

[29]  G. Ramsay,et al.  Drosophila myb exerts opposing effects on S phase, promoting proliferation and suppressing endoreduplication. , 2002, Development.

[30]  M. Botchan,et al.  Role for a Drosophila Myb-containing protein complex in site-specific DNA replication , 2002, Nature.

[31]  J. Bishop,et al.  Drosophila myb is required for the G2/M transition and maintenance of diploidy. , 1998, Genes & development.

[32]  M. Schartl,et al.  Platyfish and swordtails: a genetic system for the analysis of molecular mechanisms in tumor formation. , 1995, Trends in genetics : TIG.

[33]  S. Kamel‐Reid,et al.  Quantitative real-time PCR identifies a critical region of deletion on 22q13 related to prognosis in oral cancer , 2002, Oncogene.

[34]  W. Gerald,et al.  MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells , 2001, Nature.

[35]  T. Golub,et al.  A Mechanism of Cyclin D1 Action Encoded in the Patterns of Gene Expression in Human Cancer , 2003, Cell.

[36]  Leonard I. Zon,et al.  Cancer genetics and drug discovery in the zebrafish , 2003, Nature Reviews Cancer.