Cloning and genomic organization of beclin 1, a candidate tumor suppressor gene on chromosome 17q21.

The beclin 1 (BECN1) gene encodes a 60-kDa coiled-coil protein that interacts with the prototypic apoptosis inhibitor Bcl-2. Previous studies indicate that beclin 1 maps to a region approximately 150 kb centromeric to BRCA1 on chromosome 17q21 that is commonly deleted in breast, ovarian, and prostate cancer. The complete cDNA sequence of beclin 1 encodes a 2098-bp transcript, with a 120-bp 5' UTR, 1353-bp coding region, and 625-bp 3' UTR. Hybridization screening of a human genomic PAC library identified PAC 452O8, which contains the complete beclin 1 gene. Determination of the exon-intron structure of beclin 1 reveals 12 exons, ranging from 61 to 794 bp, which extend over 12 kb of the human genome. FISH analysis of human breast carcinoma cell lines using PAC 452O8 as probe identified allelic beclin 1 deletions in 9 of 22 cell lines. Sequencing of genomic DNA from 10 of these cell lines revealed no mutations in coding regions or splice junctions. Additionally, Northern blot analysis of 11 cell lines did not identify any abnormalities in beclin 1 transcripts. These results indicate that human breast carcinoma cell lines frequently contain allelic deletions of beclin 1, but not beclin 1 coding mutations.

[1]  D. Eccles,et al.  Early loss of heterozygosity on 17q in ovarian cancer. The Abe Ovarian Cancer Genetics Group. , 1992, Oncogene.

[2]  W. Dunn,et al.  Autophagy and related mechanisms of lysosome-mediated protein degradation. , 1994, Trends in cell biology.

[3]  Y. Nakamura,et al.  Allelotype of human ovarian cancer. , 1991, Cancer research.

[4]  P. Seglen,et al.  Reduced autophagic activity in primary rat hepatocellular carcinoma and ascites hepatoma cells. , 1993, Carcinogenesis.

[5]  K. Kinzler,et al.  Has the breast cancer gene been found? , 1994, Cell.

[6]  N. Sonenberg,et al.  Differential efficiencies of in vitro translation of mouse c-myc transcripts differing in the 5' untranslated region. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[7]  M. Soares,et al.  Construction and characterization of a normalized cDNA library. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[8]  James E. Goldman,et al.  Protection against Fatal Sindbis Virus Encephalitis by Beclin, a Novel Bcl-2-Interacting Protein , 1998, Journal of Virology.

[9]  R. Berkowitz,et al.  A 400 kb novel deletion unit centromeric to the BRCA1 gene in sporadic epithelial ovarian cancer. , 1996, Oncogene.

[10]  L. Cannon-Albright,et al.  A P1-based physical map of the region from D17S776 to D17S78 containing the breast cancer susceptibility gene BRCA1. , 1994, Human molecular genetics.

[11]  S. Kametaka,et al.  Apg14p and Apg6/Vps30p Form a Protein Complex Essential for Autophagy in the Yeast, Saccharomyces cerevisiae * , 1998, The Journal of Biological Chemistry.

[12]  R Rothstein,et al.  A high-resolution annotated physical map of the human chromosome 13q12-13 region containing the breast cancer susceptibility locus BRCA2. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[13]  M. King,et al.  Mutation analysis of the BRCA1 gene in ovarian cancers. , 1995, Cancer research.

[14]  W. Lowry,et al.  Allele loss from chromosome 17 in ovarian cancer. , 1990, Oncogene.

[15]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[16]  L. Hartmann,et al.  Human epithelial ovarian cancer allelotype. , 1993, Cancer research.

[17]  M. Kozak.  Structural features in eukaryotic mRNAs that modulate the initiation of translation. , 1991, The Journal of biological chemistry.

[18]  M. Kozak.  Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes , 1986, Cell.

[19]  M. Soares,et al.  Cloning and gene mapping of the chromosome 13q14 region deleted in chronic lymphocytic leukemia. , 1997, Genomics.

[20]  J. Uitto,et al.  Structural organization of the human type VII collagen gene (COL7A1), composed of more exons than any previously characterized gene. , 1994, Genomics.

[21]  P. Sharp,et al.  Splicing of messenger RNA precursors. , 1987, Science.

[22]  S. Emr,et al.  Endosome to Golgi Retrieval of the Vacuolar Protein Sorting Receptor, Vps10p, Requires the Function of the VPS29, VPS30, and VPS35 Gene Products , 1997, The Journal of cell biology.

[23]  D. Grignon,et al.  Loss of heterozygosity of the BRCA1 and other loci on chromosome 17q in human prostate cancer. , 1995, Cancer research.

[24]  Ming K. Lee,et al.  22 genes from chromosome 17q21: cloning, sequencing, and characterization of mutations in breast cancer families and tumors. , 1995, Genomics.

[25]  N Risch,et al.  Genetic analysis of breast cancer in the cancer and steroid hormone study. , 1991, American journal of human genetics.

[26]  James M. Roberts,et al.  The murine gene p27Kip1 is haplo-insufficient for tumour suppression , 1998, Nature.

[27]  Steven E. Bayer,et al.  A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. , 1994, Science.

[28]  J. D. Engel,et al.  An unusual 5' splice sequence is efficiently utilized in vivo. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[29]  R Rothstein,et al.  Assembly of ordered contigs of cosmids selected with YACs of human chromosome 13. , 1994, Genomics.

[30]  F. Collins,et al.  Somatic mutations in the BRCA1 gene in sporadic ovarian tumours , 1995, Nature Genetics.

[31]  J. Rommens,et al.  Generation of a transcription map at the HSD17B locus centromeric to BRCA1 at 17q21. , 1995, Genomics.

[32]  N. Dean,et al.  Serine/Threonine Protein Phosphatase Type 5 Acts Upstream of p53 to Regulate the Induction of p21WAF1/Cip1 and Mediate Growth Arrest* , 1998, The Journal of Biological Chemistry.

[33]  S. Pulst,et al.  Genomic structure of the human gene for spinocerebellar ataxia type 2 (SCA2) on chromosome 12q24.1. , 1998, Genomics.

[34]  M. Skolnick,et al.  BRCA1 mutations in primary breast and ovarian carcinomas. , 1994, Science.

[35]  Marvin B. Shapiro,et al.  RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. , 1987, Nucleic acids research.

[36]  E. Schon,et al.  High-resolution YAC-cosmid-STS map of human chromosome 13. , 1998, Genomics.

[37]  J. Barrett,et al.  Detection of frequent allelic loss on proximal chromosome 17q in sporadic breast carcinoma using microsatellite length polymorphisms. , 1992, Cancer research.

[38]  A. Knudson.  Mutation and cancer: statistical study of retinoblastoma. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[39]  Y. Nakamura,et al.  Detailed deletion mapping of chromosome 17q in ovarian and breast cancers: 2-cM region on 17q21.3 often and commonly deleted in tumors. , 1993, Cancer research.

[40]  M. Kozak,et al.  An analysis of vertebrate mRNA sequences: intimations of translational control , 1991, The Journal of cell biology.